Rh(D)-binding proteins and magnetically activated cell sorting method for production thereof

ABSTRACT

The invention includes Rh(D) binding proteins, including antibodies, and DNA encoding such proteins. Methods of generating such proteins and DNAs are also included.

CROSS REFERENCE TO RELATED APPLICATION

This application is entitled to priority pursuant to 35 U.S.C. §119(e)to U.S. provisional patent application No. 60/081,380, which was filedon Apr. 10, 1998, and is a continuation-in-part of U.S. application Ser.No. 08/884,045, filed Jun. 27, 1997, which (now U.S. Pat. No. 5,876,925)application is entitled to priority under 35 U.S.C. §119(e) to U.S.Provisional Application No. 60/028,550, filed on Oct. 11,1996.

GOVERNMENT SUPPORT

This invention was supported in part by a grant from the U.S. Government(NIH Grant No. P50-HL54516) and the U.S. Government may therefore havecertain rights in the invention.

FIELD OF THE INVENTION

The field of the invention is generation of binding proteins.

BACKGROUND OF THE INVENTION

The ability to produce monoclonal antibodies has revolutionizeddiagnostic and therapeutic medicine. Monoclonal antibodies are typicallyproduced by immortalization of antibody-producing mouse lymphocytes thusensuring an endless supply of cells which produce mouse antibodies.However, for many human applications, it is desirable to produce humanantibodies. For example, it is preferable that antibodies which areadministered to humans for either diagnostic or therapeutic purposes arehuman antibodies since administration of human antibodies to a humancircumvents potential immune reactions to the administered antibody,which reactions may negate the purpose for which the antibody wasadministered.

In addition, there exists certain situations where, for diagnosticpurposes, it is essential that human antibodies be used because otheranimals are unable to make antibodies against the antigen to be detectedin the diagnostic method. For example, in order to determine the Rhphenotype of human red blood cells (RBCs), human sera that containsanti-Rh antibody must be used since other animals cannot make anantibody capable of detecting the human Rh antigen.

The production of human antibodies in vitro by immortalizing human Blymphocytes using Epstein Barr virus (EBV)-mediated transformation orcell fusion has been fraught with technical difficulties due to therelatively low efficiency of both EBV-induced transformation and cellfusion when compared with the murine system. To overcome these problems,processes have been developed for the production of human antibodiesusing M13 bacteriophage display (Burton et al., 1994, Adv. Immunol.57:191-280). Essentially, a cDNA library is generated from mRNA obtainedfrom a population of antibody-producing cells. The mRNA encodesrearranged immunoglobulin (Ig) genes and thus, the cDNA encodes thesame. Amplified cDNA is cloned into M13 expression vectors creating alibrary of phage which express human Fab fragments on their surface.Phage which display the antibody of interest are selected by antigenbinding and are propagated in bacteria to produce soluble human Fab Ig.Thus, in contrast to conventional monoclonal antibody synthesis, thisprocedure immortalizes DNA encoding human Ig rather than cells whichexpress human Ig.

There are several difficulties associated with the generation ofantibodies using bacteriophage. For example, many proteins cannot bepurified in a non-denatured state, in that purification proceduresnecessarily involve solubilization of protein which may render someproteins permanently denatured with concomitant destruction of antigenicsites present thereon. Such proteins thus cannot be bound to a solidphase and therefore cannot be used to pan for phage bearing antibodieswhich bind to them. An example of such a protein is the human Rhantigen.

To solve the problem, a method was developed wherein intact RBCs wereused as the panning antigen (Siegel et al., 1994, Blood 83:2334-2344).However, it was discovered that since phage are inherently “sticky” andRBCs express a multitude of antigens on the cell surface, a sufficientamount of phage which do not express the appropriate antibody on thesurface also adhere to the RBCs, thus rendering the method impracticalfor isolation of phage which express antibody of desired specificity.

De Kruif et al. (1995, Proc. Natl. Acad. Sci. USA 92:3938-3942) disclosea method of isolating phage encoding antibodies, whereinantibody-expressing phage are incubated with a mixture ofantigen-expressing cells and cells which do not express antigen. Theantibody-expressing phage bind to the antigen-expressing cells.Following binding with phage, a fluorescently labeled antibody is addedspecifically to the antigen-expressing cells, which cells are removedfrom the mixture having antibody-expressing phage bound thereto. Theisolation of fluorescently labeled cells is accomplished using thetechnique of fluorescently-activated cell sorting (FACS), an expensiveand time-consuming procedure.

There remains a need for a method of isolating recombinant proteins,preferably antibodies, which is rapid and economical, and which willprovide a vast array of protein-binding proteins useful for diagnosticand therapeutic applications in humans.

SUMMARY OF THE INVENTION

The invention relates to an isolated protein having an amino acidsequence comprising a sequence selected from the group consisting of SEQID NOs: 1-69 and 139-181. In one embodiment, the isolated protein is anantigen-binding protein. In one aspect, the antigen is human Rh(D)protein. In another embodiment, the binding protein has an amino acidsequence selected from the group consisting of SEQ ID NOs: 1-69 and139-181. In one aspect, the binding protein is an antibody. In anotheraspect, the said antibody comprises a heavy chain having an amino acidsequence selected from the group consisting of SEQ ID NOs: 1-28 and139-153. In still another aspect, the antibody comprises a light chainhaving an amino acid sequence selected from the group consisting of SEQID NOs: 29-69 and 154-181. In yet another aspect, the antibody comprisesa heavy chain having an amino acid sequence selected from the groupconsisting of SEQ ID NOs: 1-28 and 139-153 and a light chain having anamino acid sequence selected from the group consisting of SEQ ID NOs:29-69 and 154-181.

In another embodiment of the isolated binding protein, the bindingprotein is an antibody fusion protein.

In another embodiment of the isolated protein, the protein issubstantially purified.

The invention also includes an isolated DNA encoding the isolatedprotein of the invention. In one embodiment, the isolated DNA has anucleotide sequence selected from the group consisting of SEQ ID NOs:70-138 and 182-224. In another embodiment, the DNA is substantiallypurified.

The invention also includes an isolated DNA encoding a protein obtainedby generating a synthetic DNA library in a virus vector expressing saidprotein; adding a magnetic label to cells expressing saidantigen-bearing moiety; incubating virus expressing said protein withsaid magnetically labeled cells in the presence of an excess ofnon-labeled cells which do not express said antigen-bearing moiety toform a mixture, wherein said virus binds to said magnetically labeledcells; isolating virus bound cells from said mixture and obtaining DNAencoding said protein therefrom. In one embodiment, the DNA has anucleotide sequence selected from the group consisting of SEQ ID NOs:70-138 and 182-224.

The invention further includes a substantially pure protein obtained bygenerating a synthetic DNA library in a virus vector expressing saidprotein; adding a magnetic label to cells expressing saidantigen-bearing moiety; incubating virus expressing said protein withsaid magnetically labeled cells in the presence of an excess ofnon-labeled cells which do not express said antigen-bearing moiety toform a mixture, wherein said virus binds to said magnetically labeledcells; isolating virus bound cells from said mixture and isolating saidprotein therefrom. In one embodiment, the protein has an amino acidsequence selected from the group consisting of SEQ ID NOs: 1-69 and139-181.

The invention also includes a substantially pure preparation of aprotein obtained by expressing said protein from DNA encoding saidprotein, wherein said DNA is obtained by generating a synthetic DNAlibrary in a virus vector expressing said protein; adding a magneticlabel to cells expressing said antigen-bearing moiety; incubating virusexpressing said protein with said magnetically labeled cells in thepresence of an excess of non-labeled cells which do not express saidantigen-bearing moiety to form a mixture, wherein said virus binds tosaid magnetically labeled cells; isolating virus bound cells from saidmixture and obtaining DNA encoding said protein therefrom. In oneembodiment, the protein has an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 1-69 and 139-181.

The invention further relates to a method of isolating a DNA encoding amulti-subunit protein which binds to an antigen-bearing moiety. Thismethod comprises

(a) generating a phage display library comprising a plurality of virusvectors. A first of the virus vectors comprises a first heterologous DNAencoding a subunit of the protein and expresses the subunit on thesurface thereof. A second of the virus vectors comprises a secondheterologous DNA encoding a different subunit of the protein andexpresses the different subunit on the surface thereof.

(b) adding a magnetic label to cells bearing the antigen-bearing moietyon their surface.

(c) incubating the phage display library with the magnetically labeledcells in the presence of an excess of non-labeled cells which do notexpress the antigen-bearing moiety to form a mixture. The first andsecond virus vectors thereby bind to the magnetically labeled cells.

(d) isolating magnetically labeled cells from the mixture. The first andsecond virus vectors are thereby isolated from the mixture.

(e) obtaining the first heterologous DNA from the first virus vector.

(f) ligating at least the portion of the first heterologous DNA encodingthe subunit and at least the portion of the second heterologous DNAencoding the different subunit to form a hybrid heterologous DNA.

(g) generating a hybrid virus vector comprising the hybrid heterologousDNA and expressing the subunit and the different subunit of the proteinon the surface thereof.

(h) adding a magnetic label to cells bearing the antigen-bearing moietyon their surface.

(i) incubating the hybrid virus vector with the magnetically labeledcells in the presence of an excess of non-labeled cells which do notexpress the antigen-bearing moiety to form a mixture. The hybrid virusvector thereby binds to the magnetically labeled cells.

(j) isolating magnetically labeled cells from the mixture. The hybridvirus vector is thereby isolated from the mixture.

(k) obtaining DNA encoding the protein from the isolated virus vector.The DNA is thereby isolated.

The invention also relates to a method of isolating a multi-subunitprotein which binds to an antigen-bearing moiety. This method comprises

(a) generating a phage display library comprising a plurality of virusvectors. A first of the virus vectors comprises a first heterologous DNAencoding a subunit of the protein and expresses the subunit on thesurface thereof. A second of the virus vectors comprises a secondheterologous DNA encoding a different subunit of the protein andexpresses the different subunit on the surface thereof.

(b) adding a magnetic label to cells bearing the antigen-bearing moietyon their surface.

(c) incubating the phage display library with the magnetically labeledcells in the presence of an excess of non-labeled cells which do notexpress the antigen-bearing moiety to form a mixture. The first andsecond virus vectors thereby bind to the magnetically labeled cells.

(d) isolating magnetically labeled cells from the mixture. The first andsecond virus vectors are thereby isolated from the mixture.

(e) obtaining the first heterologous DNA from the first virus vector.

(f) ligating at least the portion of the first heterologous DNA encodingthe subunit and at least the portion of the second heterologous DNAencoding the different subunit to form a hybrid heterologous DNA.

(g) generating a hybrid virus vector comprising the hybrid heterologousDNA and expressing the subunit and the different subunit of the proteinon the surface thereof.

(h) adding a magnetic label to cells bearing the antigen-bearing moietyon their surface.

(i) incubating the hybrid virus vector with the magnetically labeledcells in the presence of an excess of non-labeled cells which do notexpress the antigen-bearing moiety to form a mixture. The hybrid virusvector thereby binds to the magnetically labeled cells.

(j) isolating magnetically labeled cells from the mixture. The hybridvirus vector is thereby isolated from the mixture.

(k) isolating the protein from the isolated virus vector. The protein isisolated.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a diagram of a strategy for cell-surface Fab-phage panningusing magnetically-activated cell sorting.

FIG. 2 is a graph depicting cell-surface biotinylation of human RBCs.

FIG. 3 is a series of graphs which validate the antigen-positive,antigen-negative cell separation procedure of the invention.

FIG. 4 is an image of a microplate agglutination assay whereinanti-Rh(D) Fab/phage agglutination titer was measured.

FIG. 5 is an image of a microplate agglutination assay showingdetermination of Rh(D) binding epitope for selected anti-Rh(D) Fab/phageclones.

FIG. 6 is an image depicting the use of Fab/phage antibodies in a gelcard assay.

FIG. 7 comprises FIGS. 7A and 7B. FIG. 7A is a dendrogram which depictsthe relationship among the anti-Rh(D) heavy chains described herein inExample 2. The 28 unique heavy chain clones are organized by V_(H)family, V_(H) germline gene, and VDJ rearrangement. Each heavy chainclone is identified by a numeral preceded by a letter (“B” through “E”)which denotes its gerinline gene. The 28 heavy chains comprised 12distinct VDJ regions, designated VDJ1-VDJ12. Clones with identical VDJjoins putatively result from intra-clonal diversity of 12 original Blymphocytes. FIG. 7B is an alignment of the CDR3 regions of theanti-Rh(D) heavy chains.

FIG. 8 comprises FIGS. 8A, 8B, and 8C. FIG. 8A, comprising FIGS. 8A and8A-1 through 8A-4, is an alignment of anti-Rh(D) heavy chains to theirnearest germline V, D, and J genes. Also illustrated are the putativeintermediate heavy chain sequences (Ca, Cb, Da, Db, Dc). The number ofnucleotide differences from a germline V_(H) is tabulated to the rightof each sequence. In general, D segments showed poor homology with knownD genes so mutations were not scored in these regions. Replacementmutations are indicated with letters, silent mutations are indicated as“*”, identities are indicated as “.”, and insertions are indicated as“-”. Sequences derived from the 5′V_(H) primers used in libraryconstruction are indicated as “>”. FIG. 8A is a map demonstrating howFIGS. 8A-1 to 8A-4 are arranged. FIG. 8B, comprising FIGS. 8B, 8B-1, and8B-2 is an alignment of the four VH3 genes utilized by anti-Rh(D) heavychains. FIG. 8B is a map depicting the arrangement of FIGS. 8B-1 and8B-2. FIG. 8C is a dendrogram which depicts the relationship among humanVH3 family germline genes, and illustrate relatedness of VH3-21, VH3-30.VH3-33, and VH3-30.3 and the surprising restriction in V_(H) gene usage.The VH3-30.5 gene is present in only certain haplotypes and is identicalto VH3-30.

FIG. 9 is an ontogenic tree of anti-Rh(D) heavy chains constructed usingnucleotide alignment data. Circles represent isolated and sequencedclones, and diamonds represent putative intermediates. The number ofnucleotide mutations from its germline V_(H) gene is indicated inparentheses below the clone name. The distance along the horizontal axisrepresents the degree of mutation (including J segments) within theconstraints of the diagram.

FIG. 10 comprises FIGS 10A and 10B. FIG. 10A, comprising FIGS. 10A and10A-1 through 10A-4, is an alignment of anti-Rh(D)κ light chains totheir nearest germline V and J genes, and indicates predominance ofDPK-9 usage from the V_(κ)I family. Nomenclature for clones is similarto that for heavy chains but uses the letters “F” through “I”. FIG. 10Ais a map depicting the arangement of FIGS. 10A-1 through 10a-4. FIG.10B, comprising FIGS. 10B, 10B-1, and 10B-2 is an alignment of the fourV_(κ) genes utilized by anti-Rh(D) light chains. FIG. 10B is a mapshowing the arrangement of FIGS. 10B-1 and 10B-2. Symbols are the sameas those used in FIG. 8A.

FIG. 11 comprises FIG. 11A and 11B. FIG. 11A comprising FIGS. 11A and11A-1 through 11A-4, is an alignment of anti-Rh(D)λ light chains totheir nearest germline V and J genes. FIG. 11A is a map depicting thearrangement of FIGS. 11A-1 through 11A-4. FIG. 11B, comprising FIGS.11B, 11B-1, and 11B-2, is an alignment of the 10V_(λ) germline genesutilized, and illustrates the use of a diverse set of variable regiongenes derived from multiple families. However, all of the clones use theidentical J_(λ) gene segment. FIG. 11B is a map representing thearrangement of FIGS. 11B-1 and 11B-2. Nomenclature for the clones issimilar to that for heavy chains but uses the letters “J” through “S”.Symbols are the same as those used in FIG. 8A.

FIG. 12, comprising FIGS. 12A, 12B, and 12C, is a trio of graphs whichdepict comparisons of variable region gene family usage foranti-Rh(D)-specific clones and randomly-picked, non-Rh(D)-binding clonesfrom original γ₁κ and γ₁λ non-selected libraries. Upwardly-angledhatched bars reveal heterogeneity in V_(H) (FIG. 12A), V_(κ) (FIG. 12B),and V_(λ) (FIG. 12C) family representation before selection foranti-Rh(D) specificity. Numbers above bars represent absolute number ofclones in that group.

FIG. 13 depicts the results of determinations of the Rh(D) bindingepitope of anti-Rh(D) Fab/phage clones. The five different agglutinationpatterns obtained from screening all of the 53 Fab/phage clones areillustrated. The particular clones shown in FIG. 13 are identified bytheir unique heavy chain/light chain pairings using the nomenclaturedefined in FIGS. 7, 10, and 11. For E1/M3, reactivity with additionalRh(D) variant cells is required to distinguish its specificity for epD3from that for epD9. Inclusion of the category IVb cell permits theidentification of a new epitope designated “epDX”.

FIG. 14, comprising FIGS. 14A and 14B, is matrix illustrating thegenetic composition and epitope specificity of anti-Rh(D) antibodies.The horizontal axis represents the unique γ₁ heavy chains and thevertical axis represents the unique (FIG. 14A) and (FIG. 14B) λ and κlight chains (based on amino acid sequence). A shaded pattern at theintersection of a heavy chain/light chain pair indicates the Rh(D)epitope specificity observed for that Fab/phage antibody. A few clonesgave mixed patterns of reactivity as described herein. Although heavychains D1, D15, D16, and D17 differ in nucleotide sequence, these chainshave an identical amino acid sequence and thus comprise a single column.Similarly, heavy chains C5 and C8 and λ light chains K1 and K2 encodethe same proteins. The pairings of these 28 heavy and 41 light chainnucleotide gene segments, which produced 53 unique Fab transcripts,encoded 43 different Fab proteins, as indicated in the matrix.

FIG. 15, comprising FIGS. 15A, 15B, and 15C, depicts the results ofinhibition studies performed using recombinant anti-Rh(D) antibodies.The figures show results of representative experiments demonstrating themutual inhibition of antibodies directed at two different Rh(D) epitopes(in this example, epD3 and epD6/7, FIGS. 15A and 15C), but not betweenan Rh(D) antibody and an unrelated recombinant anti-RBC antibody (ananti-blood group B antibody, FIG. 15B). In FIG. 15A, Rh(D)-positive RBCswere incubated with soluble Fabs only, phage-displayed Fabs only, orcombinations of the two, as indicated. In FIG. 15B, Rh(D)-positive RBCsthat were blood group B were used. After washing, RBCs were resuspendedin anti-M13 antibody and assessed for agglutination induced byphage-displayed Fabs. Soluble Fabs were used “full-strength” whileFab/phage preparations were present in limiting amounts to increase thesensitivity of the inhibition assay, as described herein. In FIG. 15C,mutual inhibition of epD3 and epD6/7 anti-Rh(D) antibodies wasdemonstrated with Rh(D)-positive RBCs, γ₁κ and γ₁λ soluble Fabs, andlight chain isotype-specific antisera (see text for details). In theseexamples, the anti-epD3 and anti-epD6/7 antibodies were clones E1/M3 andD5/I3, respectively. The anti-blood group B antibody was isolated froman IgG phage display library made from the splenic B cells of a bloodgroup O donor.

FIG. 16, comprising FIGS. 16A, 16B, and 16C, depict models for Rh(D)antigen/antibody binding. A conventional model (depicted in FIG. 16A)and a model described herein (depicted in FIG. 16B) for Rh(D)antigen/antibody binding predict different combining sites and geneticrelationships between antibodies. As depicted in FIG. 16C, if antibodiesdirected at different Rh(D) epitopes are clonally related, then theexpressed repertoire will differ between Rh(D)-negative and partialRh(D) individuals.

DETAILED DESCRIPTION

According to the present invention, there is provided a novel method ofisolating DNA encoding a protein and the protein encoded thereby,wherein the protein is preferably an antibody, which protein is capableof specifically binding to an antigen-bearing moiety.

As exemplified herein but not limited thereto, the method comprisesgenerating bacteriophage which encode human antibodies. Specifically inthe present invention, anti-Rh(D) RBC Fab/phage antibodies encoded by anM13 filamentous phage library are obtained. The library is generatedfrom antibody-producing cells obtained from a hyperimmunized donor byfirst obtaining cDNA derived from mRNA expressed in theantibody-producing cells. Ig encoding fragments of the cDNA are obtainedusing the polymerase chain reaction (PCR) and primers specific for suchfragments of DNA. Ig-specific DNA so obtained is cloned into abacteriophage. Bacteriophage encoding the Ig fragments are pannedagainst a mixture of antigen-positive, biotinylated RBC-target cellspre-coated with streptavidin-conjugated magnetic microbeads and excessnon-labeled RBCs. Bacteriophage which express antibodies on the phagesurface, which antibodies are specific for the target cell antigen, bindto the labeled cells. These phage are separated from phage which arebound to non-labeled cells and from phage which are not bound to thecells using a magnetic column. Phage so separated encode and displayantibody specific for antigens on the target cells.

To generate a phage antibody library, a cDNA library is first obtainedfrom mRNA which is isolated from cells which express the desired proteinto be expressed on the phage surface, e.g., the desired antibody. cDNAcopies of the mRNA are produced using reverse transcriptase. cDNA whichspecifies Ig fragments are obtained by PCR and the resulting DNA iscloned into a suitable bacteriophage vector to generate a bacteriophageDNA library comprising DNA specifying Ig genes. The procedures formaking a bacteriophage library comprising heterologous DNA are wellknown in the art and are described, for example, in Sambrook et al.(1989, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor,N.Y.).

A bacteriophage library may also be obtained using cDNA rather thanPCR-amplified Ig encoding fragments of cDNA. Generation of a cDNAlibrary is useful for the isolation of proteins which are notantibodies, such as ligands and the like.

Bacteriophage which encode the desired protein, e.g., an antibody, maybe engineered such that the protein is displayed on the surface thereofin such a manner that it is available for binding to its correspondingbinding protein, e.g., the antigen against which the antibody isdirected. Thus, when bacteriophage which express a specific antibody areincubated in the presence of a cell which expresses the correspondingantigen, the bacteriophage will bind to the cell. Bacteriophage which donot express the antibody will not bind to the cell.

For panning of bacteriophage, i.e., selection of phage which express thedesired antibody, cells which express the corresponding antigen arelabeled with a detectable label such as biotin. Streptavidin-conjugatedmagnetic beads are then added to the cells. The cells are mixed with anexcess of non-labeled cells which do not express the antigen. This cellmixture is then incubated with the phage library, wherein phage whichexpress the antibody bind to cells expressing the antigen. The presenceof the excess non-labeled cells in the mixture serves as a means ofremoving bacteriophage which do not express the antibody but which mightotherwise bind to antigen-expressing cells non-specifically. The detailsof the experimental procedures for practicing the present invention areprovided herein in the experimental detail section.

Antigen-expressing cells having antibody-expressing phage bound theretoare magnetically removed from the mixture. One example of magneticremoval involves pouring the mixture of magnetic and non-magnetic cellsinto a column in the selective presence or absence of a magnetic fieldsurrounding the column. Alternatively, magnetic cells may be separatedfrom non-magnetic cells in solution by simply holding a magnet againstthe side of a test tube and attracting the cells to the inner wall andthen carefully removing the non-magnetic cells from the solution.

Thus, the method of the invention involves a procedure for enriching apopulation of recombinant phage for those expressing specificphage-displayed ligands derived from natural or synthetic phage DNAlibraries by simultaneously performing negative and positive selectionagainst a mixture of magnetically-labeled receptor-positive particles(i.e., cells) and non-labeled receptor-negative particles.

The terms “bacteriophage” and “phage” are used interchangeably hereinand refer to viruses which infect bacteria. By the use of the terms“bacteriophage library” or “phage library” as used herein, is meant apopulation of bacterial viruses comprising heterologous DNA, i.e., DNAwhich is not naturally encoded by the bacterial virus.

The term “virus vector” includes a virus into which heterologous DNA hasbeen inserted. The virus vector may be a bacteriophage or may be aeukaryotic virus.

By the term “target cell” as used herein, is meant a cell whichexpresses an antigen against which the desired antibody is sought.

By the term “panning” or “panned” as used herein, is meant the processof selecting phage which encode the desired antibody.

By the term “Fab/phage” as used herein, is meant a phage particle whichexpresses the Fab portion of an antibody.

By the term “scFv/phage” are used herein, is meant a phage particlewhich expresses the Fv portion of an antibody as a single chain.

By “excess non-labeled cells” is meant an amount of non-labeled cellswhich exceeds the number of labeled cells. Preferably, the ratio oflabeled cells to non-labeled cells is about 1:2. More preferably, theratio of labeled cells to non-labeled cells is greater than about 1:4.Even more preferably, the ratio of labeled cells to non-labeled cells isgreater than about 1:10.

While the method of the invention as exemplified herein describes thegeneration of phage which encode the Fab portion of an antibodymolecule, the method should not be construed to be limited solely to thegeneration of phage encoding Fab antibodies. Rather, phage which encodesingle chain antibodies (scFV/phage antibody libraries) are alsoincluded in the invention. Fab molecules comprise the entire Ig lightchain, that is, they comprise both the variable and constant region ofthe light chain, but include only the variable region and first constantregion domain (CH1) of the heavy chain. Single chain antibody moleculescomprise a single chain of protein comprising the Ig Fv fragment. An IgFv fragment includes only the variable regions of the heavy and lightchains of the antibody, having no constant region contained therein.Phage libraries comprising scFV DNA may be generated following theprocedures described in Marks et al., 1991, J. Mol. Biol. 222:581-597.Panning of phage so generated for the isolation of a desired antibody isconducted as described herein for phage libraries comprising Fab DNA.

The invention should also be construed to include synthetic phagedisplay libraries in which the heavy and light chain variable regionsmay be synthesized such that they include nearly all possiblespecificities. Therefore, antibody-displaying libraries can be “natural”or “synthetic” (Barbas, 1995, Nature Medicine 1:837-839; de Kruif et al.1995, J. Mol. Biol. 248:97-105). Antibody-displaying librariescomprising “natural” antibodies are generated as described in theexperimental example section. Antibody-displaying libraries comprising“synthetic” antibodies are generated following the procedure describedin Barbas (1995, supra) and the references cited therein.

The method of the invention should be further construed to includegeneration of phage display libraries comprising phage other than M13 asexemplified herein. Other bacteriophage, such as lambda phage, may alsobe useful in the method of the invention. Lambda phage display librarieshave been generated which display peptides encoded by heterologous DNAon their surface (Sternberg et al., 1995, Proc. Natl. Acad. Sci. USA92:1609-1613). Moreover, it is contemplated that the method of theinvention may be extended to include viruses other than bacteriophage,such as eukaryotic viruses. In fact, eukaryotic viruses may be generatedwhich encode genes suitable for delivery to a mammal and which encodeand display an antibody capable of targeting a specific cell type ortissue into which the gene is to be delivered. For example, retroviralvectors have been generated which display functional antibody fragments(Russell et al., 1993, Nucl. Acids Res. 21:1081-1085).

The red blood cell antibodies to which antibodies may be generatedinclude, but are not limited to, Rh antigens, including Rh(D), Rh(C),Rh(c), Rh(E), Rh(e), and other non-Rh antigens, including red blood cellantigens in the Kell, Duffy, Lutheran and Kidd blood groups.

Thus, the method of the invention is not limited solely to the isolationof DNA encoding anti-Rh(D) antibodies, but rather may be used for theisolation of DNA encoding antibodies directed against any RBC antigen orother cell antigen, such as, but not limited to, tumor-specific antigen,bacterial antigens, and the like. The method of the invention is alsouseful for typing platelets by generating phage antibodies specific fora number of clinically important platelet antigens, notably,P1^(A1)/P1^(A2), Bak^(a)/Bak^(b), Pen^(A)/Pen^(B), and the like.

The invention is further useful for typing donor white blood cells forHLA antigens for the purposes of matching donors and recipients forpotential transplant matching in the case of both solid (for example,kidney, heart, liver, lung) and non-solid (for example, bone marrow)organ or tissue transplanting.

To detect binding of phage expressing antibody directed against one ofthese non-red blood cell antigens, the non-red blood cells may beagglutinated or trapped following the procedures described herein foragglutination or trapping of red blood cells. Prior to agglutination ortrapping, the cells may be rendered “visible” by staining or otherlabeling technique in order that agglutination or trapping is apparentto the naked eye or scanner.

The method of the invention is most useful for the generation of aprotein which binds to an antigen-bearing moiety, where theantigen-bearing moiety is not easily purified in soluble form. Thus, theantigen-bearing moiety includes antigens which are associated with otherstructures, usually membranes in the cell such as cell membranes or cellorganelle membranes.

In accordance with the present invention, the antigen-bearing moiety maybe a protein, a lipid, a carbohydrate or a nucleic acid, or it may be acomplex of at least two of a protein, a lipid, a carbohydrate and anucleic acid, it being appreciated that many antigen-bearing moieties incells are not comprised of one of these components alone. Preferably,the antigen-bearing moiety is a membrane bound protein, such as anantigen or a receptor protein. However, when the antigen-bearing moietyis a carbohydrate, it may be a carbohydrate expressed on a glycolipid,for example, a P blood group antigen or other antigen.

By the term “antigen-bearing moiety” as used herein, is meant a moleculeto which an antibody binds.

By the term “antigen-binding protein” as used herein, is meant apolypeptide molecule, such a an antibody, a fragment thereof or anantibody fusion protein, which is capable of specifically binding toanother molecule.

By the term “antibody fusion protein” as used herein, is meant apolypeptide molecule having an amino acid sequence which comprises theamino acid sequence of a portion of an antigen-binding protein. Theportion of the antigen-binding protein may, for example, be an entireantibody or a fragment thereof.

The method of the invention is also useful for the generation ofautoimmune antibodies such as those involved in autoimmune hemolyticanemia (AIHA) (Siegel et al., 1994, Structural analysis of red cellautoantibodies, Garratty (ed.) Immunobiology of Transfusion Medicine,Dekker, New York, N.Y.). Autoimmune antibodies that are directed againstcell antigens which are cell surface membrane associated or cellorganelle membrane associated may be isolated using the technologydescribed herein. Autoimmune diseases and their associated antigens towhich antibodies may be isolated include, but are not limited to thefollowing: Myasthenia gravis (acetylcholine receptor; neurons), chronicinflammatory demyelinating polyneuropathy (myelin; neurons), autoimmunethyroid disease (thyroid stimulating hormone receptor; thyroid cells),primary biliary cirrhosis (mitochondrial autoantigens; livermitochondria), idiopathic thrombocytopenic purpura (platelet membraneintegrins; platelets), pemphigus vulgaris (epidermal antigens;epidermis), and Goodpasture's syndrome (basement membrane antigens;kidney or lung cells).

In fact, the method of the invention is useful for the isolation of DNAclones encoding any antibody directed against an antigen expressed on acell, which cell can be labeled with a magnetic label and which cell canbe obtained in sufficient quantities in an non-labeled form so as toprovide an excess of non-labeled cells as required in the assay.

Further, the method of the invention is not limited to the isolation ofDNA encoding antibodies but rather may also be used for the isolation ofDNA encoding other peptides or proteins having specificity for cellproteins, such as, for example, but not limited to, ligands which bindcell receptor proteins, peptide hormones, and the like.

The invention should also not be construed as being limited to the useof biotin as the cell-labeling agent. Other labels may be used providedtheir addition to a cell does not disturb the structural integrity ofany surface proteins expressed thereon and provided such labels permitthe addition of a paramagnetic microbead or other magnetic substancethereto. Other such labels include, but are not limited to, cell surfaceproteins or carbohydrates which can be directly derivitized withmagnetic beads that possess activated amine, carboxyl, or thiol groups.In addition, dyes such as fluorescein or rhodamine may also becovalently attached to cells in a manner similar to biotin and magneticbeads coated with anti-dye antibodies may be attached thereto.

The invention also includes a screening method which may be used toisolate a DNA encoding a multi-subunit protein which binds to anantigen-bearing moiety or, alternately, to isolate the multi-subunitprotein itself. The multi-subunit protein may, for example, be anantibody or another immunoglobulin. It is well known that antibodies andother immunoglobulins comprise multiple subunits, often designated heavyand light chains.

According to this screening method, a phage display library isgenerated, either as described herein or using other generally known orhereafter-developed methods. The library comprises a plurality of virusvectors, including a first virus vector which comprises a firstheterologous DNA encoding a subunit of the protein. The first virusvector expresses the subunit on its surface, either by itself or inassociation with one or more other subunits of the protein. The libraryalso comprises a second virus vector which comprises a secondheterologous DNA encoding a different subunit of the protein. The secondvirus vector expresses the different subunit on its surface, either byitself or in association with one or more other subunits of the protein.A magnetic label is added to cells bearing the antigen-bearing moiety ontheir surface, and the labeled cells are incubated with the phagedisplay library in the presence of an excess of non-labeled cells whichdo not express the antigen-bearing moiety. The first and second virusvectors bind to the magnetically labeled cells, owing to interaction(s)between the antigen and the subunits of the protein expressed on thesurface of the vectors.

After incubating the phage display library with the mixture of cells,magnetically labeled cells are isolated from the mixture. First andsecond virus vectors bound to the magnetically labeled cells are therebyalso isolated from the mixture. The virus vectors are separated from themagnetically labeled cells (e.g. by culturing the cells in a manner inwhich the virus vectors are produced in the culture supernatant), andheterologous DNA is obtained from virus vectors that adhered to themagnetically labeled cells. The DNA may optionally be purified at thisstage. DNA isolated from the virus vectors that adhered to themagnetically labeled cells includes the first heterologous DNA and thesecond heterologous DNA.

At least the portion of the first heterologous DNA encoding the subunitis ligated to at least the portion of the second heterologous DNAencoding the different subunit to form a hybrid heterologous DNA. Forthis purpose, it is advantageous that the virus vector be constructed insuch a way that the portion of the first heterologous DNA encoding thesubunit, the portion of the second heterologous DNA encoding thedifferent subunit, or both, are flanked or surrounded by definedrestriction endonuclease cleavage sites. In such constructs, the portionof the first heterologous DNA encoding the subunit may be removed, forexample, by treating the first heterologous DNA with restrictionendonucleases which specifically cleave the specific sites. This portionmay then be ligated, for example either directly or after ligating alinker DNA thereto, to all or a portion of the second heterologous DNAto generate the hybrid heterologous DNA.

The hybrid heterologous DNA is then used to generate a hybrid virusvector comprising the hybrid heterologous DNA. The hybrid virus vectorexpresses the subunit and the different subunit of the protein on itssurface. For example, if the first heterologous DNA encodes an antibodylight chain and the second heterologous DNA encodes an antibody heavychain, then the hybrid virus vector may express an antibody comprisingequal numbers of heavy and light chains on its surface.

The hybrid virus vector is then incubated with the mixture ofmagnetically labeled cells having the antigen-bearing moiety on theirsurface and non-magnetically labeled cells which do not have theantigen-bearing moiety on their surface. Owing to interactions betweenthe antigen and the subunits of the protein expressed on the surface ofthe hybrid virus vector, the hybrid virus vector binds with themagnetically labeled cells, and may therefore be isolated from themixture of cells by isolating magnetically labeled cells from themixture.

As described herein, hybrid virus vector particles are isolated from themagnetically labeled cells. The isolated hybrid virus vectors may beused as a source for obtaining either the multi-subunit protein or thehybrid heterologous DNA (which encodes the subunits of the protein),using standard methods.

The invention includes proteins and DNA encoding the same which aregenerated using the methods described herein. To isolate DNA encoding anantibody, for example, DNA is extracted from antibody expressing phageobtained according to the methods of the invention. Such extractiontechniques are well known in the art and are described, for example, inSambrook et al. (supra).

The invention includes a number of isolated or substantially purifiedDNAs encoding antigen-binding proteins, such as Rh(D)-binding proteins.For example, a DNA having a nucleotide sequence comprising at least oneof SEQ ID NOs: 70-138 and 182-224, as described herein, is included. Theisolated or substantially purified nucleic acid may have a nucleotidesequence selected from the group consisting of SEQ ID NOs: 70-138 and182-224.

An “isolated DNA”, as used herein, refers to a DNA sequence, segment, orfragment which has been purified from the sequences which flank it in anaturally occurring state, e.g., a DNA fragment which has been removedfrom the sequences which are normally adjacent to the fragment, e.g.,the sequences adjacent to the fragment in a genome in which it naturallyoccurs. The term also applies to DNA which has been substantiallypurified from other components which naturally accompany the DNA, e.g.,RNA or DNA or proteins which naturally accompany it in the cell.

The invention also includes a number of isolated or substantiallypurified proteins, such as Rh(D)-binding proteins. For example, aprotein having an amino acid sequence comprising at least one of SEQ IDNOs: 1-69 and 139-181, as described herein, is included. The isolated orsubstantially purified protein may have an amino acid sequence selectedfrom the group consisting of SEQ ID NOs: 1-69 and 139-181. The proteinmay be an antigen-binding protein, such as an antibody which comprises aheavy chain having an amino acid sequence selected from the groupconsisting of SEQ ID NOs: 1-28 and 139-153, a light chain having anamino acid sequence selected from the group consisting of SEQ ID NOs29-69 and 154-181, or both. The protein may also be, for example, anantibody fusion protein.

An “isolated protein” as used herein, means a protein or polypeptidewhich has been separated from components which naturally accompany it ina cell. Typically, a protein or polypeptide is isolated when at least10%, more preferably at least 20%, more preferably at least 50% of thetotal material (by volume, by wet or dry weight, or by mole percent ormole fraction) in a sample is the protein or polypeptide of interest.

The invention should also be construed to include DNAs which aresubstantially homologous to the DNA isolated according to the method ofthe invention. Preferably, DNA which is substantially homologous isabout 50% homologous, more preferably about 70% homologous, even morepreferably about 80% homologous and most preferably about 90% homologousto DNA obtained using the method of the invention.

“Homologous” as used herein, refers to the subunit sequence similaritybetween two polymeric molecules, e.g., between two nucleic acidmolecules, e.g., two DNA molecules or two RNA molecules, or between twopolypeptide molecules. When a subunit position in both of the twomolecules is occupied by the same monomeric subunit, e.g., if a positionin each of two DNA molecules is occupied by adenine, then they arehomologous at that position. The homology between two sequences is adirect function of the number of matching or homologous positions, e.g.,if half (e.g., five positions in a polymer ten subunits in length) ofthe positions in two compound sequences are homologous then the twosequences are 50% homologous, if 90% of the positions, e.g, 9 of 10, arematched or homologous, the two sequences share 90% homology. By way ofexample, the DNA sequences 3′ATTGCC 5′ and 3′TATGCG 5′ share 50%homology.

To obtain a substantially pure preparation of a protein comprising, forexample, an antibody, generated using the methods of the invention, theprotein may be extracted from the surface of the phage on which it isexpressed. The procedures for such extraction are well known to those inthe art of protein purification. Alternatively, a substantially purepreparation of a protein comprising, for example, an antibody, may beobtained by cloning an isolated DNA encoding the antibody into anexpression vector and expressing the protein therefrom. Protein soexpressed may be obtained using ordinary protein purification procedureswell known in the art.

As used herein, the term “substantially pure” describes a compound,e.g., a protein or polypeptide which has been separated from componentswhich naturally accompany it. Typically, a compound is substantiallypure when at least 10%, more preferably at least 20%, more preferably atleast 50%, more preferably at least 60%, more preferably at least 75%,more preferably at least 90%, and most preferably at least 99% of thetotal material (by volume, by wet or dry weight, or by mole percent ormole fraction) in a sample is the compound of interest. Purity can bemeasured by any appropriate method, e.g., in the case of polypeptides bycolumn chromatography, gel electrophoresis or HPLC analysis. A compound,e.g., a protein, is also substantially purified when it is essentiallyfree of naturally associated components or when it is separated from thenative contaminants which accompany it in its natural state.

As used herein, amino acids are represented by the full name thereof, bythe three letter code corresponding thereto, or by the one-letter codecorresponding thereto, as indicated in the following table:

Full Name Three-Letter Code One-Letter Code Aspartic Acid Asp D GlutamicAcid Glu E Lysine Lys K Arginine Arg R Histidine His H Tyrosine Tyr YCysteine Cys C Asparagine Asn N Glutamine Gln Q Serine Ser S ThreonineThr T Glycine Gly G Alanine Ala A Valine Val V Leucine Leu L IsoleucineIle I Methionine Met M Proline Pro P Phenylalanine Phe F Tryptophan TrpW

The present invention also provides for analogs of proteins or peptidesobtained according to the methods of the invention. Analogs can differfrom naturally occurring proteins or peptides by conservative amino acidsequence differences or by modifications which do not affect sequence,or by both.

For example, conservative amino acid changes may be made, which althoughthey alter the primary sequence of the protein or peptide, do notnormally alter its function. Conservative amino acid substitutionstypically include substitutions within the following groups:

glycine, alanine;

valine, isoleucine, leucine;

aspartic acid, glutamic acid;

asparagine, glutamine;

serine, threonine;

lysine, arginine;

phenylalanine, tyrosine.

Modifications (which do not normally alter primary sequence) include invivo, or in vitro chemical derivatization of polypeptides, e.g.,acetylation, or carboxylation. Also included are modifications ofglycosylation, e.g., those made by modifying the glycosylation patternsof a polypeptide during its synthesis and processing or in furtherprocessing steps; e.g., by exposing the polypeptide to enzymes whichaffect glycosylation, e.g., mammalian glycosylating or deglycosylatingenzymes. Also embraced are sequences which have phosphorylated aminoacid residues, e.g., phosphotyrosine, phosphoserine, orphosphothreonine.

Also included in the invention are polypeptides which have been modifiedusing ordinary molecular biological techniques so as to improve theirresistance to proteolytic degradation or to optimize solubilityproperties. Analogs of such polypeptides include those containingresidues other than naturally occurring L-amino acids, e.g., D-aminoacids or non-naturally occurring synthetic amino acids. The peptides ofthe invention are not limited to products of any of the specificexemplary processes listed herein.

In addition to substantially full length polypeptides, the presentinvention provides for active fragments of the polypeptides. A specificpolypeptide is considered to be active if it binds to an antigen-bearingmoiety, for example, if a fragment of an antibody binds to itscorresponding antigen in the same manner as the full length protein.

As used herein, the term “fragment,” as applied to a polypeptide, willordinarily be at least about fifty contiguous amino acids, typically atleast about one hundred contiguous amino acids, more typically at leastabout two hundred continuous amino acids and usually at least aboutthree hundred contiguous amino acids in length.

The invention is further described in detail by reference to thefollowing experimental examples. These examples are provided forpurposes of illustration only, and are not intended to be limitingunless otherwise specified. Thus, the invention should in no way beconstrued as being limited to the following examples, but rather, shouldbe construed to encompass any and all variations which become evident asa result of the teaching provided herein.

EXAMPLE 1 Isolation of Cell Surface-Specific Human Monoclonal AntibodiesUsing Phage Display and Magnetically-Activated Cell Sorting

The experiments described in this Example provide procedures and resultsfor the isolation and production of anti-Rh(D) red blood cell antibodiesusing Fab/phage display.

A method is described in FIG. 1 for the isolation of filamentousphage-displayed human monoclonal antibodies specific for non-purifiablecell surface expressed molecules. To optimize the capture ofantigen-specific phage and minimize the binding of irrelevant phageantibodies, a simultaneous positive and negative selection strategy wasemployed. Cells bearing the antigen of interest are pre-coated withmagnetic beads and are diluted into an excess of unmodifiedantigen-negative cells. Following incubation of the cell admixture witha Fab/phage library, the antigen positive cell population is retrievedusing magnetically-activated cell sorting, and antigen-specificFab/phage are eluted and propagated in bacterial culture. When thisprotocol was used with magnetically-labeled (Rh(D)-positive and excessnon-labeled Rh(D)-negative human red blood cells and a Fab/phage libraryconstructed from human peripheral blood lymphocytes, dozens of unique,clinically useful _(γ1)κ and _(γ1)λ anti-Rh(D) antibodies were isolatedfrom a single alloimmunized individual.

The cell-surface selection method of the present invention is readilyadaptable for use in other systems, such as for the identification ofputative tumor-specific antigens, and provides a rapid (less than onemonth), high yield approach for isolating self-replicative antibodyreagents directed at novel or conformationally-dependent cell-surfaceepitopes.

Creation of Fab/phage Display Libraries

Separate _(γ1)κ and _(γ1)λ phage libraries were constructed from 2×10⁷mononuclear cells derived from the peripheral blood from anRh(D)-negative individual previously hyperimmunized with Rh(D)-positivered blood cells (RBCs). The phagemid vector pComb3 (Barbas, 1991, Proc.Natl. Acad. Sci. USA 88:7978-7982) was used to create the librariesutilizing previously published methods (Barbas et al., 1991,Combinatorial immunoglobulin libraries on the surface of phage (Phabs):Rapid selection of antigen-specific Fabs. Methods: A Companion toMethods in Enzymology 2:119-124; Siegel et al., 1994, Blood83:2334-2344).

Briefly, cDNA was prepared from the mRNA of the donor cells and heavychain and light chain immunoglobulin (Ig) cDNA segments were amplifiedusing the polymerase chain reaction (PCR) and the battery of human Igprimers described by Kang et al. (1991, “Combinatorial ImmunoglobulinLibraries on the Surface of Phage (Phabs): Rapid Selection ofAntigen-Specific Fabs. Methods: A Companion to Methods” in Enzymology2:111-118) supplemented by those of Silverman et al. (1995, J. Clin.Invest. 96:417-426). The heavy and light chain PCR products were clonedinto pComb3 and electroporated into E. coli. Upon co-infection withVCSM13 helper phage (Stratagene, La Jolla, Calif.), Ig DNA was packagedinto filamentous phage particles which express human Fab molecules fusedto the gene III bacteriophage coat protein.

Panning Fab Phage Display Libraries for Anti-Rh(D) Clones

Rh(D)-positive RBCs were cell-surfaced biotinylated by incubating cellsat a hematocrit of 10% with 500 μg/ml sulfo-NHS-LC-biotin (PierceChemical, Rockford, Ill.) for 40 minutes at room temperature (RT).Following 5 washes with phosphate-buffered saline (PBS), 8×10⁶biotinylated Rh(D)-positive RBCs were incubated with 10 μl ofstreptavidin-coated paramagnetic microbeads (MACS StreptavidinMicrobeads, Mitenyi Biotec, Sunnyvale, Calif.) for 1 hour at RT in atotal volume of 100 μl PBS. Non-reacted beads were removed by washingand then the magnetic bead-coated, Rh(D)-positive RBCs were mixed with a10-fold excess (8×10⁷) of the Rh(D)-negative (unmodified) RBCs and˜3×10¹¹ colony-forming units (cfu) of either the _(γ1)κ and _(γ1)λFab/phage libraries (prepared as described above) in a final volume of40 μl PBS containing 2% non-fat dry milk (MPBS, Carnation, Nestle FoodProducts, Glendale, Calif.).

Following a 2 hour incubation at 37° C., the RBC/phage suspension wasloaded at a flow rate of 10 μl/minute onto a MiniMACS magnetic type MScolumn (Mitenyi Biotec, Sunnyvale, Calif.) that was pre-equilibratedwith 2% MPBS. This loading step was performed without a magnetic fieldaround the column so as to prevent magnetic bead-coated RBCs frominstantly adhering to the very top of the column, clogging it, andcausing the trapping of Rh(D)negative non-biotinylated RBCs. Loading theRBC/phage incubation mixture in the absence of a magnetic field causesthe antigen-negative and antigen-positive RBCs to distribute evenlythroughout the column without running off since the excluded volume ofthe column is slightly greater than 40 μl. Once loaded, the column wasplaced in a magnetic field (MiniMACS magnetic separation unit, MitenyiBiotec, Sunnyvale, Calif.) for 2 minutes to allow the Rh(D)-positiveRBCs to adhere, and a series of 500 μl washes were performed withice-cold MPBS followed by a final wash with PBS. A total of 3 washeswere performed for the first 2 rounds of panning and a total of 6 washeswere performed for all subsequent pannings. For each panning, the firstwash was carried out at a flow rate of 10 μl/minute during which timethe bulk of Rh(D)-negative RBCs washed off the column. All subsequentwashes were performed at 200 μl/minute. Following the last wash, thecolumn was removed from the magnetic field and thebead-coated/phage-coated Rh(D)-positive RBCs were flushed off the columnwith 500 μl PBS using the plunger from a 5 cc syringe (Becton-Dickinson,Franklin Lakes, N.J.).

The RBCs were immediately centrifuged for 5 seconds at 13,000×g and werethen resuspended in 200 μl of 76 mM citrate, pH 2.4, to denature theRh(D) antigen and elute bound phage. Following a 10 minute incubationperiod at RT with intermittent vortexing, the phage eluate and cellulardebris were neutralized with 18 μl 2 M Tris base and were added to 10 mlof O.D.=1.0 XL1-Blue strain of E. coli (Stratagene, La Jolla, Calif.)grown in super broth (SB) (Barbas et al., 1991, supra) supplemented with10 μg/ml tetracycline. After incubation for 15 minutes at RT, duringwhich time the phage library enriched for Rh(D) binders was allowed toinfect the bacterial culture, 10 ml of pre-warmed, 37° C. SB containing40 μg/ml carbenicillin/10 μg/ml tetracycline was added to give finalantibiotic concentrations of 20 μg/ml and 10 μg/ml, respectively. Asmall aliquot of culture (˜100 μl) was immediately removed and titeredon Luria broth/carbenicillin plates to determine the number of phagecontained in the total eluate. The balance of the culture was shaken at37° C. for 1 hour at 300 RPM. Additional antibiotics, additional SB, andVCSM13 helper phage were subsequently added and the culture was grownovernight at 30° C. as described (Siegel et al., 1994, supra).

Phagemid particles were purified from the culture supernatant bypolyethylene glycol 8000 (PEG) precipitation (Barbas et al., 1991,supra), resuspended in 1% bovine serum albumin (BSA)/PBS, and dialyzedovernight to remove residual PEG that may lyse RBCs during subsequentrounds of panning. Thus, the resultant phage preparation serves as theinput for the next round of panning. The _(γ1)κ and _(γ1)λ phagelibraries were panned separately to prevent any bias in light chainisotype replication possibly introduced by bacterial amplification.

Screening Polyclonal Fab/Phage Libraries and Individual Phage Coloniesfor Anti-Rh(D) Reactivity

The specificity of Fab/phage for the Rh(D) antigen was assessed usinganti-M13 antibody as a bridging antibody to induce agglutination betweenRBCs that have bound anti-Rh(D) Fab/phage. One hundred μl aliquots ofpolyclonal Fab/phage from rounds of panning, or monoclonal Fab/phagederived from individual Fab/phage eluate clones, were incubated with 50μl of a 3% suspension of RBCs of defined phenotype (i.e., Rh(D)-negativeor -positive).

Following 1 hour incubation at 37° C., the RBCS were washed 3 times with2 ml cold PBS to remove unbound Fab/phage. The resultant RBC pelletswere resuspended in 100 μl of a 10 μg/ml solution of sheep anti-M13antibody (5-Prime 3-Prime, Boulder, Col.) and transferred to theround-bottomed wells of a 96-well microtiter plate. Plates were leftundisturbed (˜2 hours) and were then read. Wells having a negativereaction exhibit sharp ˜2 millimeter diameter RBC spots whereas in wellshaving positive reactions, i.e., agglutination, the RBCs in agglutinatedwells form a thin carpet coating the entire floor of the well.

For hemagglutination assays utilizing mini-column gel cards(ID-Micro-Typing System, Ortho Diagnostics, Raritan, N.J.) (Lapierre etal., 1990, Transfusion 30:109-113), 25 μl of Fab/phage clones were mixedwith 50 μl aliquots of RBCs (0.8% suspensions in Micro Typing Systembuffer, Ortho Diagnostics). The mixtures were placed in the reservoirsabove the mini-columns which contain dextran-acrylamide beads previouslysuspended in 100 μl/ml anti-M13 antibody. After incubation at 37° C.,the gel cards were centrifuged at 70×g for 10 minutes and were read.

Miscellaneous Methods

Preparation of fluorescently-labeled RBCs for flow cytometry wasperformed as described herein and samples were analyzed using a FACScanmicrofluorimeter equipped with Lysis II (Ver 1.1) software(Becton-Dickinson, Mountain View, Calif.). Plasmid DNA was prepared frombacterial clones (Qiawell Plus, Qiagen, Chatsworth, Calif.).Double-stranded DNA was sequenced using light chain or heavy chain Igconstant region reverse primers or unique pComb3 vector primers thatanneal 5-prime to the respective Ig chain (Barbas et al., 1991, supra;Roben et al., 1995, J. Immunol. 154:6437-6445) and automatedfluorescence sequencing (Applied Biosystems, Foster City, Calif.).Sequences were analyzed using MacVector Version 5.0 sequencing software(Oxford Molecular Group, Oxford, UK) and the Tomlinson database of Iggermline genes (Tomlinson et al., 1996, V Base Sequence Directory. MRCCenter for Protein Engineering, Cambridge, UK).

Experimental Design for Cell Incubation and Separation Protocols

The experimental conditions described above for panning Fab/phagelibraries for anti-RBC-reactive phage were determined after performing aseries of initial studies aimed at optimizing the cell separationprocess and ultimate yield of antigen-specific Fab/phage. The mainparameters investigated included:

Biotinylation Conditions were sought that would biotinylate the RBCsurface in a manner such that a sufficient number of streptavidin-coatedmagnetic beads would bind to the cells causing the RBCs to be retainedby a magnetic column. In this case, over-biotinylation that mightdestroy the antigenicity of the Rh(D) antigen or might make the cellsnon-specifically absorb antibody is to be avoided. To address thisissue, Rh(D)-positive/Kell-negative RBCs (Kell being a RBC antigen;(Walker, ed. 1993, In: Technical Manual, 11^(th) Ed., Bethesda, Md.,American Association of Blood Banks) were incubated with a range ofsulfo-NHS-LC-biotin concentrations and the degree of biotinylation wasassessed by flow cytometry utilizing fluorescein-conjugatedstreptavidin.

To assess the degree of cell-surface biotinylation, 5 μl aliquots of 3%suspensions of Rh(D)-positive/Kell-negative RBCs biotinylated at varyingbiotin reagent concentrations were incubated with 200 μl of a 1/100dilution of FITC-streptavidin (Jackson ImmunoResearch, Bar Harbor,Maine) for 30 min at 4° C. (FIG. 2). The mixture was washed withphosphate buffered saline (PBS) and analyzed by flow microfluorimetry(-□-). Aliquots of cells were also analyzed for retention ofRh(D)-antigenicity (-Δ-) (i.e., specific staining) or for lack ofnon-specific staining (-◯-) by incubating the cells with 100 μl ofeither anti-Rh(D) or anti-Kell typing sera, respectively, washing thecells and then staining them with a 1/100 dilution of FITC-goatanti-human IgG (Jackson ImmunoResearch).

A linear, non-saturating response was observed (FIG. 2). Retention ofRh(D) antigenicity was assessed using anti-Rh(D) typing serum and wasfound to be unaffected by the derivatization of cell-surface proteinswith biotin at all biotin concentrations tested (FIG. 2). Furthermore,the Kell-negative RBCs did not non-specifically adsorb anti-Kellantibodies.

Each biotinylated RBC sample was then incubated with an excess ofstreptavidin-coated magnetic microbeads and applied to a magneticseparation column. It was determined that as many as 10⁸ RBCs could beretained by the column for RBC samples biotinylated with greater than orequal to 500 μg/ml biotin reagent. Since the actual RBC/phage panningexperiments were designed to use only ˜10⁷ Rh(D)-positive cells (seebelow), RBC biotinylation at 500 μg/ml was determined to be sufficient.

Concentration of Rh(D)-Positive and Rh(D)-Negative RBCs in IncubationMixture

Prior to performing Fab/phage panning experiments, the ability of themagnetically-activated cell separation technique to separateRh(D)-positive and Rh(D)-negative cells was assessed using anti-Rh(D)typing serum and flow cytometry (FIG. 3). Streptavidin-microbead coated,biotinylated Rh(D)-positive RBCs (8×10⁶ cells) were mixed with a 10-foldexcess of Rh(D)-negative non-coated RBCs (8×10⁷ cells) in a 40 μl volumeof PBS containing 2% non-fat dry milk (MPBS) and the mixture was appliedto a MiniMACS column. The column was washed and the bound cells wereeluted as described herein. Aliquots of RBCs contained in the originaladmixture (panel a), the column wash (panel b), and the column eluate(panel c) were stained with anti-Rh(D) typing serum and FITC-goatanti-human IgG as described in FIG. 2. The flow cytograms show thatalthough ˜90% of the cells in the column load were Rh(D)-negative (panela), nearly all of them washed off of the column (panel b), yielding acolumn eluate that was almost entirely Rh(D)-positive cells (panel c).Since only ˜6% of the final eluate comprise Rh(D)-negative cells (panelc), and Rh(D)-negative cells were initially present in a 10-fold excessto Rh(D)-positive cells, only ˜0.6% of the initial antigen-negativeimmunosorbent cells contaminated the final antigen-positive preparation.This efficiency of the cell separation was deemed adequate forsubsequent panning experiments with Fab/phage.

In the above-described experiment, to avoid clogging the magneticseparation column, it was necessary to load the column in the absence ofa magnetic field. This necessitated a reaction volume of less than orequal to 40 μl so that none of the material would run off the column. Ontheoretical grounds (Kretzschmar et al., 1995, Anal. Biochem.224:413-419), one can calculate the appropriate concentration of cellsrequired in a 40 μl volume to capture greater than 50% of Fab/phagespecific for a given cell surface antigen. Such a calculation is afunction of the number of antigen sites per cell and the dissociationconstant (K_(D)) of the bound Fab/phage. Using a value of ˜100,000 Rh(D)antigen sites per RBC (phenotype “—D—/—D—”) (Mollison et al., 1993, In:Blood Transfusion in Clinical Medicine, Oxford, Blackwell ScientificPublications) and the desired Fab/phage affinity in the K_(D)=10⁻⁸ to10⁻⁹ M range, then 8×10⁶ Rh(D)-positive RBCs in a 40 μl reaction volumewould be required. Given this number of Rh(D)-positive cells, a 10-foldexcess of Rh(D)-negative RBCs was found to be the maximum amount ofantigen-negative cells that could be effectively separated fromantigen-positive RBCs by the magnetic column (FIG. 3).

Construction and Panning of Fab/phage Libraries

_(γ1)κ and _(γ1)λ phage libraries were prepared as described herein andwere found to contain 7×10⁷ and 3×10⁸ independent transformants,respectively. Table 1 tabulates the panning results for the libraries.

An RBC agglutination assay utilizing anti-M13 secondary antibody asbridging antibody was used to detect anti-Rh(D) Fab/phage activity inthe panned polyclonal libraries and the individual randomly-pickedFab/phage clones (FIG. 4). The results shown are a representativeexample of the assay depicting negative reactivity to Rh(D)-negativeRBCs and strongly positive reactivity to Rh(D)-positive RBCs for the_(γ1)κ library (panning #2) out to a dilution of 1/2048.

In the case of the _(γ1)κ library, significant enrichment for bindingphage appears to occur after only one round of panning, whereassignificant enrichment for the _(γ1)λ library occurs during the secondround. This is reflected by both the sharp increase in the percent ofphage bound during a given round of panning as well as the ability ofthe polyclonal _(γ1)κ and _(γ1)λ Fab/phage libraries to agglutinateRh(D)-positive RBCs after 1 and 2 rounds of panning, respectively (Table1, FIG. 4).

Monoclonal Fab/phage were prepared from randomly-picked individualbacterial colonies obtained during each round of panning. It wasapparent that by the third round of panning, all clones have anti-Rh(D)specificity (Table 1). To confirm that these Fab/phage have anti-Rh(D)specificity and are not binding to other unrelated antigens that maycoincidentally be present on the particular Rh(D)-positive RBC andabsent on the particular Rh(D)-negative RBC used in the agglutinationassays, clones were screened against a panel of 11 Rh(D)-negativeand-positive RBCs of varying blood group specificities to verify theiranti-Rh(D) specificity (Walker, 1993, supra).

Clonal Analysis at the Genetic Level

To investigate the genetic diversity among the randomly pickedanti-Rh(D) clones, plasmid DNA was prepared from each of the clones andthe corresponding heavy and light chain Ig nucleotide sequences wereidentified. In Table 2 there is listed a number of attributes for eachclone including the name of the most closely-related germline heavy orlight chain Ig gene. More detailed analysis at the nucleotide levelrevealed that among all of the anti-Rh(D) binding clones, there were alarge number of unique heavy and light chain DNA sequences (Table 3).Because of the random shuffling of heavy and light chain gene segmentswhich occurs during the creation of a Fab/phage display library (Barbaset al., 1991, supra), it is evident that these heavy chains and lightchains combined to form nearly 50 different anti-Rh(D) antibodies.

A detailed multiple alignment analysis of the predicted amino acidsequences revealed a total of twenty-five unique heavy chain, eighteenunique kappa light chain and twenty-three unique lambda light chainproteins. Due to the combinatorial effect during library construction,these heavy and light chain gene segments paired to produce fifty uniqueFab antibodies (20_(γ1κ) and 30_(γ1λ)). Of interest, all twenty fiveunique heavy chains and nearly all of the eighteen unique kappa lightchains were derived from only 5V_(H)III or four VκI germline genes,respectively, while the lambda light chains were derived from a morediverse set of germline genes. Analysis of heavy and light chainnucleotide sequences from over sixty negative clones from the non-pannedlibraries were performed to verify the heterogeneity in variable regionfamily representation before selection. Clones representing V_(H)families I (13%), III (36%), IV (31%), V(15%) and VI (5%); Vκ families I(43%), II (14%), III (29%) and IV (14%); and Vγ families I (48%), II(4%), III (9%), IV (4%), V (9%), VI (17%) and VII (9%) were present.

Clonal Analysis at the Protein Level

To investigate the diversity in fine specificity (Rh(D) antigen epitopespecificity) among the anti-Rh(D) clones, agglutination experiments wereperformed with selected clones and with sets of rare Rh(D)-positive RBCswhich were obtained from individuals whose RBCs produce Rh(D) antigenlacking certain epitopes. Examining the pattern of agglutination of aparticular anti-Rh(D) antibody with such sets of mutant RBCs enables theidentification of the specific epitope on Rh(D) to which the antibody isdirected (Mollison et al., 1993, supra). A representative example ofsuch an experiment is shown in FIG. 5 and the Rh(D) epitopes forselected anti-Rh(D) Fab/phage clones are tabulated in Table 2.

Agglutination experiments were performed with anti-Rh(D)-negative RBCs(rr), Rh(D)-positive RBCs (R₂R₂), and “partial” Rh(D)-positive RBCs(mosaics IIIa, IVa, Va, VI, VII). The results shown are a representativeexample of the assay for 5 randomly-picked anti-Rh(D) Fab/phage clones(FIG. 5).

TABLE 1a _(γ1)κFAB/PHAGE LIBRARY PANNING RESULTS AGGLU φINPUT φOUTPUT %BOUND⁴ T BINDERS/ PANNING¹ (CFUs)² (CFUs)³ (× 10⁻⁴) ENRICHMENT⁵ TITER⁶TOTAL (%)⁷ 0 0 0/16 (0) 1 2.94 × 10¹¹ 6.04 × 10⁵ 2.1 1/16  0/16 (0) 22.15 × 10¹¹ 1.68 × 10⁷ 78.3 38.0 x 1/2048 15/15 (100) 3 1.72 × 10¹¹ 1.44× 10⁸ 840.0 10.7 x 1/2048 12/12 (100)

TABLE 1a _(γ1)κFAB/PHAGE LIBRARY PANNING RESULTS AGGLU φINPUT φOUTPUT %BOUND⁴ T BINDERS/ PANNING¹ (CFUs)² (CFUs)³ (× 10⁻⁴) ENRICHMENT⁵ TITER⁶TOTAL (%)⁷ 0 0 0/16 (0) 1 2.94 × 10¹¹ 6.04 × 10⁵ 2.1 1/16  0/16 (0) 22.15 × 10¹¹ 1.68 × 10⁷ 78.3 38.0 x 1/2048 15/15 (100) 3 1.72 × 10¹¹ 1.44× 10⁸ 840.0 10.7 x 1/2048 12/12 (100)

TABLE 2a ANALYSIS OF _(γ1)κFAB/PHAGE CLONES AGGL VH V_(κ) CLONE¹ U² FAM³VH GENE⁴ FAM⁵ V_(ρ) GENE⁶ D EPITOPE⁷ KPO-1 neg 3 DP-47/V3-23 4DPK24/VklVKlobeck KPO-2 neg 3 DP-31/V3-9P 3 DPK22/A27 KPO-3 neg 3DP-58/hv3d1EG 4 DPK24/VklVKlobeck KPO-4 neg 4 3d279d+ — no light chainKPO-5 neg 3 DP-29/12-2 1 LFVK431 KPO-6 neg 4 DP-79/4d154 1 DPK9/012KPO-7 neg 3 V3-48/hv3d1 4 DPK24/VklVKlobeck KPO-8 neg 4 DP-70/4d68 2DPK18/A17 KPO-9 neg 1 DP-14/V1-18 1 DPK9/012 KPO-10 neg 4 DP-70/4d68 1DPK9/012 KPO-11 neg 5 DP-73/V5-51 1 DPK9/012 KPO-12 neg 3 DP-54/V3-7 2DPK18/A17 KPO-13 neg 3 V3-48/hv3d1 1 Vb′ KPO-14 neg 6 DP-74/VH-VI 1DPK6/Vb″ KPO-15 neg 3 DP-46/3d216 3 Vg/38K KPO-16 neg 6 DP-74/VH-VI 1DPK9/012 KP1-1 neg 4 V71-4+ 3 DPK22/A27 KP1-2 neg 4 3d279d+ 1 DPK8/Vd+KP1-3 neg 1 4M28 1 DPK9/012 KP1-4 neg 4 DP-79/4d154 3 Vg/38K KP1-5 neg 3DP-38/9-1 3 DPK22/A27 KP1-6 neg 4 DP-70/4d68 1 L12a/PCRdil6-5 KP1-7 neg5 DP-73/V5-51 2 DPK15/A19 KP1-8 neg 4 DP-70/4d68 3 DPK22/A27 KP1-9 neg —no heavy chain — no light chain KP1-10 neg — no heavy chain 3 DPK22/A27KP1-11 neg 1 DP-15/V1-8+ 1 DPK9/012 KP1-12 neg 3 b28e — no light chainKP1-13 neg 3 DP-47/V3-23 4 DPK24/VklVKlobeck KP1-14 neg 3 DP-31/V3-9P 3DPK21/humkv328h5 KP1-15 neg 1 DP-7/21-2 4 DPK24/VklVKlobeck KP1-16 neg 5DP-73/V51 3 DPK22/A27 KP2-1 pos 3 DP-50/hv3019b9 1 DPK9/012 epD6/7 KP2-2pos 3 DP-50/hv3019b9 1 DPK9/012 epD6/7 KP2-3 pos 3 DP-50/hv3019b9 1DPK9/012 epD6/7 KP2-4 pos 3 b28m 1 DPK9/012 epD2 KP2-5 pos 3 b28m 1DPK9/012 epD1 KP2-6 pos 3 DP-50/hv3019b9 1 DPK9/012 epD6/7 KP2-7 pos 3DP-50/hv3019b9 1 DPK9/012 epD5 KP2-8 pos 3 DP-50/hv3019b9 1 DPK9/012KP2-9 pos 3 DP-50/hv3019b9 1 DPK9/012 epD2 KP2-10 pos 3 DP-50/hv3019b9 1DPK9/012 epD2 KP2-11 pos 3 DP-50/hv3019b9 1 DPK9/012 epD2 KP2-12 pos 3DP-50/hv3019b9 1 DPK9/012 epD1 KP2-13 pos 3 DP-50/hv3019b9 1 DPK9/012epD6/7 KP2-14 pos 3 DP-50/hv3019b9 2 DPK15/A19 epD2 KP2-15 pos 3DP-50/hv3019b9 1 DPK9/012 epD6/7 KP3-1 pos 3 DP-50/hv3019b9 1 DPK9/012KP3-2 pos 3 DP-50/hv3019b9 1 DPK9/012 epD6/7 KP3-3 pos 3 DP-50/hv3019b91 DPK9/012 KP3-4 pos 3 DP-49/1.9111 1 DPK9/012 epD5 KP3-5 pos 3DP-50/hv3019b9 1 DPK9/012 KP3-6 pos 3 DP-50/hv3019b9 1 A30/SG3+ epD6/7KP3-7 pos 3 DP-50/hv3019b9 1 DPK8/Vd+ epD6/7 KP3-8 pos 3 DP-50/hv3019b91 DPK9/012 epD6/7 KP3-9 pos 3 DP-50/hv3019b9 1 DPK9/012 KP3-10 pos 3DP-50/hv3019b9 1 DPK9/012 KP3-11 pos 3 DP-50/hv3019b9 1 DPK9/012 KP3-12pos 3 DP-46/3d216 1 DPK9/012 ¹nomenclature: prefix “KPO” denotes“_(γ1)κFab/phage library, panning 0”, “KP1” denotes “_(γ1)κFab/phagelibrary, panning 1”, etc. ²agglutination negative or positive againstRh(D)-positive RBC ³Ig heavy chain variable region gene family perTomlinson et al., supra ⁴closest related Ig heavy chain variable regiongene per Tomlinson et al. supra ⁵Ig light chain variable region genefamily per Tomlinson et al., supra ⁶closest related Ig light chainvariable region gene per Tomlinson et al., supra ⁷Rh(D) epitope asdefined by rare RBC agglutination pattern (see FIG. 5 and text)

TABLE 2b ANALYSIS OF _(τ1)ρFAB/PHAGE CLONES VH V_(ρ) CLONE¹ U² FAM³ VHGENE⁴ FAM⁵ V_(κ) GENE⁶ D EPITOPE⁷ LPO-1 neg 4 DP-65/3d75d 1 DPL7/IGLV1S2LPO-4 neg 4 DP-70/4d68 6 IGLV8A1 LPO-3 neg 6 DP-74/VH-VI 7 DPL18/VL7.1LPO-4 neg 3 DP-29/12-2 1 DPL3/Iv122 LPO-5 neg 3 DP-38/9-1 6IGLV6S1/LV6SW-G LPO-6 neg 1 4M28 1 DPL3/Iv122 LPO-7 neg 1 8M27 1DPL2/Iv1L1 LPO-8 neg 5 DP-58/V5-51 6 IGLV6S1/LV6SW-G LPO-9 neg 5DP-73/V5-51 1 DPL7/1GLV1S2 LPO-10 neg 3 DP-38/9-1 1 DPL2/Iv1L1 LPO-11neg 3 DP-31/V3-9P 3 DPL23/VLIII.1 LPO-12 neg — no heavy chain 1DPL7/1GLV1S2 LPO-13 neg 3 DP-47/V3-23 — no light chain LPO-14 neg 4DP-71/3d197d 6 IGLV6S1/LV6SW-G LPO-15 neg 4 DP-70/4d68 4 IGLV8A1 LPO-16neg 3 DP-54/V3-7 7 DPL19 LP2-1 pos 3 DP-50/hv3019b9 1 DPL2/Iv1L1 epD2LP2-2 pos 3 DP-77/WHG16 1 DPL3/Iv122 LP2-3 pos 3 DP-49/1.9111 1DPL3/Iv122 epD1 LP2-4 neg 4 3d279d+ 1 DPL2/Iv1L1 LP2-5 pos 3DP-49/1.9111 3 DPL16/IGLV3S1 epD5 LP2-6 pos 3 DP-50/hv3019b9 1DPL7/IGLV1S2 epd2 LP2-7 pos 3 b28m 1 DPL7/IGLV1S2 epD2 LP2-8 pos 3DP-49/1.9111 3 IGLV3S2=Iv318 epD1 LP2-9 pos 3 DP-50/hv3019b9 3DPL16/IGLV3S1 epD2 LP2-10 pos 3 DP-77/WHG16 1 DPL3/LV122 LP2-11 neg 1DP-75-VI-2 1 DPL5/LV117d LP2-12 pos 3 DP-77/WHG16 1 DPL2/LV1L1 epD2LP2-13 pos 3 COS-8/hv3005f3 4 IGLV8A1 LP2-14 pos 3 DP-49/1.9111 1DPL7/IGLV1S2 epD5 LP2-15 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 LP2-16 pos3 DP-49/1.9111 2 Iv2046 epd1 LP2-17 pos 3 DP-77/WHG16=V3-21+ 1DPL3/Iv122 epD3/9 LP2-18 pos 3 DP-49/1.9111 2 VL2.1˜DPL10/Iv2066 epD1LP2-19 pos 3 DP-50/hv3019b9 3 DPL16/1GLV3S1 epD2 LP2-20 neg 3 V3-49+ 3DPL16/1GLV3S1 LP2-21 pos 3 DP-50/hv3019b9 7 DPLIS/VL7.1 epD6/7 LP2-22pos 3 DP-49/1.9111 2 Iv2046 LP2-23 pos 3 DP-49/1.9111 3 DPL16/IGLV3S1epD5 LP2-24 pos 3 DP-77/WHG16 1 DPL3/Iv122 LP2-25 pos 3 DP-50/v3019b9 7DPL18/VL7.1 epD6/7 LP2-26 pos 3 DP-49/1.9111 3 DPL16/IGLV3S1 LP2-27 neg3 COS-6/DA-8 2 VL2.1 LP2-28 pos 3 COS-8/hv3005f3 4 IGLV8A1 LP2-29 pos 3DP-49/1.9111 DPL13 LP2-30 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 LP2-31pos 3 DP-50/hv3019b9 7 DPL18/VL7.1 LP2-32 pos 3 DP-49/1.9111 1DPL2/Iv1L1 LP2-33 pos 3 DP-50/hv3019b9 7 DPL18/VL7.1 LP2-34 pos 3DP-50/hv3019b9 7 DPL18/VL7.1 LP2-35 pos 3 DP-50/hv3019b9 3 DPL16/1GLV3S1LP2-36 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 LP3-1 pos 3 DP-50/hv3019b9 3DPL16/IGLV3S1 epD2 LP3-2 pos 3 DP-49/1.9111 3 DPL16/IGLV3S1 epD1 LP3-3pos 3 DP-49/1.9111 3 DPL16/IGLV3S1 LP3-4 pos 3 DP-50/hv3019b9 7DPL18/VL7.1 epD6/7 LP3-5 pos 3 DP-49/1.9111 1 DPL5/LV117d epD5 LP3-6 pos3 DP-49/1.9111 1 DPL5/LV117d epD1 LP3-7 pos 3 DP-77/WHG16 1 DPL2/Iv1L1epD5 LP3-8 pos 3 b28m 1 DPL7/IGLVIS2 epD2 LP3-9 pos 3 DP-50/hv3019b9 3DPL16/IGLV3S1 epD2 LP3-10 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 LP3-11pos 3 DP-50/hv3019b9 3 DPLI6/IGLV3S1 epD2 LP3-12 pos 3 COS-8/hv3005f3 4IGLV8A1 epD6/7 LP3-13 pos 3 DP-50/hv3019b9 1 DPL2/Iv1L1 epD2 LP3-14 pos3 DP-49/1.9111 3 DPL16/IGLV3S1 LP3-15 pos 3 DP-77/WHG16 1 DPL3/Iv122epD1 LP3-16 pos 3 DP-49/1.9111 1 DPL2/Iv1L1 epD5 LP3-17 pos 3DP-50/hv3019b9 3 DPL16/IGLV3S1 LP3-18 pos 3 DP-50/hv3019b9 3DPL16/IGLV3S1 LP3-19 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 epD5 LP3-20pos 3 DP-50/hv3019b9 1 DPL2/Iv1L1 LP3-21 pos 3 DP-49/1.911 1 DPL3/Iv122LP3-22 pos 3 COS-8/hv3005f3 1 DPL2/Iv1L1 LP3-23 pos 3 DP-49/1.9111 3DPL16/IGLV3S1 LP3-24 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 ¹nomenclature:prefix “LPO” denotes “_(γ1)λFab/phage library, panning 0”, “LP1” denotes“_(γ1)λFab/phage library, panning 1”, etc. ²agglutination negative orpositive against Rh(D)-positive RBC ³Ig heavy chain variable region genefamily per Tomlinson et al., supra ⁴closest related Ig heavy chainvariable region gene per Tomlinson et al. supra ⁵Ig light chain variableregion gene family per Tomlinson et al., supra ⁶closest related Ig lightchain variable region gene per Tomlinson et al., supra ⁷Rh(D) epitope asdefined by rare RBC agglutination pattern (see FIG. 5 and text)

TABLE 3 SUMMARY OF FAB/PHAGE CLONAL ANALYSIS Number of unique heavychains 25 Number of unique κ light chains 18 Number of unique λ lightchains 23 Number of _(γ1)κ antibodies 20 Number of _(γ1)λ antibodies 30Number Rh(D) epitope specificities represented  5

Use of Fab/Phage Antibodies as Blood Bank Typing Reagents

The ability of the anti-Rh(D) Fab/phage preparations to accuratelydistinguish Rh(D)-negative from Rh(D)-positive RBCs in microplatehemagglutination assays (FIGS. 4 and 5) provided evidence that a geltest (Lapierre et al., 1990, Transfusion 30:109-1130) used by bloodbanks to phenotype RBCs using conventional antisera could be adapted foruse with Fab/phage.

The gel test comprises a plastic card of approximately 5×7 cm,containing 6 mini-columns each filed with about 20 μl ofdextran-acrylamide beads suspended in anti-human globulin (Coombsreagent). Red cells to be typed are incubated with the desired humananti-sera and are centrifuged through the gel. RBCs which are positivefor antigens to which the antisera is directed agglutinate as theyencounter the anti-human globulin and become trapped in or above the gelmatrix. Unreactive RBCs sediment through the gel particles and form apellet at the bottom of the microtube. Because the gel test offers anumber of advantages over traditional blood banking methods for RBCphenotyping including decreased reagent volumes, the elimination of acell washing step and a more objective interpretation of results, manyblood bank facilities have adapted this new technology. As shown in FIG.6, anti-Rh-(D) Fab/phage can be used with gel cards that are modified tocontain anti-M13 antibody.

To perform the assay, Rh(D)-negative or -positive red blood cells wereincubated with dilutions of anti-Rh(D) Fab/phage (_(γ1)κ library,panning #2) and were centrifuged into micro-columns containing beadssuspended in anti-M13 antibody. Undiluted Fab/phage stock had a titer of5×10¹² cfu/ml similar to that in the microplate settling assay (FIG. 4).Because the volume of Fab/phage used in this assay is one-fourth of thatin the microplate assay, the amount of Fab/phage present in the 1/625dilution is approximately equal to that present in the 1/2048 dilutionin FIG. 4. Therefore, the number of Fab/phage required to yield apositive result is essentially equivalent in both assays.

In other assays which were performed as just described, when anti-M13antibody was eliminated from the assay, no agglutination of red bloodcells was observed. In addition, anti-IgG antibody does not react withrecombinant Fabs expressed on the surface of the bacteriophage. OnlyRh-positive cells which were reacted with anti-Rh phage wereagglutinated when anti-M13 antibody was present in the assay. It shouldbe noted that when high concentrations of anti-M13 antibody were used,even Rh-negative cells appeared to be agglutinated. This is an artifactresulting from the cross-linking of unbound (i.e., non-reacted) phagewhich becomes crosslinked in the presence of high amounts of anti-M13antibody and forms a semi-impenetrable mat through which not all theRh-negative cells can traverse. In the experiments described herein, ananti-M13 concentration of about 100 μg/ml was considered to be optimalfor agglutination and for the prevention of false positive results.Depending on the precise concentrations of reagents and cells used inthe assay, the concentration of anti-M13 may deviate from this number.

To assess the relative sensitivity of an anti-M13 modified Micro TypingSystem, the columns of the Micro Typing System cards had added to them100 μg/ml of anti-M13 antibody. Rh-negative or Rh-positive red bloodcells were incubated with undiluted or with five-fold serial dilutions(1/5, 1/25, 1/125, 1/625 and 1/3125) of anti-Rh phage antibodies. Thecards were centrifuged and samples were assessed for agglutination. Themodified Micro Typing System card assay was capable of detecting anti-Rhagglutination at a dilution of between 1/625 and 1/3125.

Procedures for Isolation of Tumor-Specific Antibodies

Fab/phage specific for tumor cells are useful for in vitro diagnosis(lab assays of biopsy, fluid, or blood samples), in vivo labeling oftumor/metastasis (coupling of antibody to imaging probe), or fortreatment of malignancy (coupling of antibodies to chemical orradioactive toxins). Tumor-specific antibodies are also useful for theidentification of novel antigens or markers on tumor cells which mayform the basis for anti-tumor vaccines. Further, tumor-specificantibodies useful for the generation of anti-idiotypic antibodies mayalso form the basis for anti-tumor vaccines.

Anti-tumor antibodies are generated essentially as described herein forthe generation of anti-Rh antibodies. Tumor cells, for example, but notlimited to, malignant melanoma cells, are cell-surface biotinylated,labeled with streptavidin-magnetic microbeads, and are then mixed withexcess normal melanocytes. Fab/phage libraries are generated fromperipheral blood lymphocytes of melanoma patients who possesstherapeutically useful anti-tumor antibodies. A number of melanomapatients who have “cured” themselves apparently have done so by mountinga humoral (ie., antibody) immune response. These Fab/phage libraries areincubated with the admixture of cells. Fab/phage which are directedagainst epitopes specific for malignant cells will bind to the malignantcells and may then be isolated utilizing the magnetic column panningapproach.

Isolation of Fab/Phage that Identify Bacterial Virulence Factors

The approach described herein may be used to isolate Fab/phage capableof detecting differences between the virulent bacteria and theirnonpathogenic counterparts. In this case, the virulent strain ofbacteria is magnetically labeled, diluted with the non-pathogeniccounterpart, and an Fab/phage library which is generated fromlymphocytes obtained from individuals infected with the virulent strainis added. Fab/phage which are isolated in this manner may be useful forthe identification of novel bacterial antigens against whichantibacterial compounds and/or vaccines may be developed.

EXAMPLE 2 Genetic and Immunological Properties of Phage-Displayed HumanAnti-Rh(D) Antibodies

Clinically, the human Rh(D) antigen is the most important red blood cell(RBC) membrane protein in transfusion medicine. The alloimmune responseagainst Rh(D) produces high affinity IgG antibodies which causehemolytic transfusion reactions and hemolytic disease of the newborn(HDN). The prophylactic use of Rh(D)-immune globulin in pregnantRh(D)-negative women has been a major advance in the prevention of HDN,yet the mechanism by which the drug exerts its immune modulatory effectis not well understood.

Monoclonal antibodies derived from the B cells of Rh(D)-immune globulindonors have defined several dozen Rh(D) epitopes (Scott, 1996, Transfus.Clin. Biol. 3:333). Paradoxically, the Rh(D) antigen, a circa 30 kDtransmembrane protein, has minimal extracellular mass and presents avery limited surface area for epitope expression. Because molecularcloning of a large repertoire of anti-Rh(D) antibodies has notpreviously been performed, these observations remain non-reconciled.

Rational development of recombinant formulations of Rh(D)-immuneglobulin would be facilitated by molecular cloning of a large number ofanti-Rh(D) antibodies. Such cloning would also aid in the design oftherapeutic agents that block antibody binding. Furthermore,comprehensive genetic analysis of anti-Rh(D) antibodies within a givenalloimmunized individual would serve as a paradigm for human immunerepertoire development, an area of which limited information iscurrently available. Previously, no more than 8 IgG anti-Rh(D) humanmonoclonal antibodies have been derived from a single individual(Boucher et al., 1997, Blood 89:3277).

In Example 1, a technique useful for isolating Fab/phage antibodiesdirected against antigens expressed on cell surfaces was described.Using this technique and intact human red blood cells (RBCs), highlydiverse γ₁κ and γ₁λ. Fab/phage libraries against the Rh(D) antigen fromthe B cells of a single Rh(D)-immune globulin donor were generated.

In this Example, a detailed genetic and serological analysis of 53unique anti-Rh(D) antibodies derived from 83 randomly chosen clones ispresented. These data demonstrate extensive genetic homology betweenantibodies directed against different Rh(D) epitopes. Evidence isprovided herein that antibodies directed against different epitopes canbe clonally related. Finally, a model is described which reconciles theserological diversity of anti-Rh(D) antibodies with the topologicalconstraints imposed by the Rh(D) antigen.

The materials and methods used in the experiments presented in thisExample are now described.

Production of Monoclonal Anti-Rh(D) Phage-Displayed and Soluble FabMolecules

Methods for the isolation of human anti-Rh(D)-specific antibodies fromγ₁κ and γ₁λ Fab/phage display libraries using the pComb3H phagemidvector and a cell-surface panning protocol have been described (Siegelet al., 1997, J. Immunol. Meth. 206:73). Soluble anti-Rh(D) Fabpreparations for inhibition studies were produced from bacterialcultures transfected with plasmid DNA from which the M13 gene III coatprotein sequence had been excised as described (Siegel et al., 1994,Blood 83:2334; Barbas et al., 1991, Methods: A Companion to Meth.Enzymol. 2:119). Cultures were grown by shaking at 300 RPM at 37° C. insuperbroth (30 g/L tryptone, 20 g/L yeast, 10 g/L MOPS, pH 7.00)containing 20 mM MgCl₂ and 50 mg/ml carbenicillin to an OD₆₀₀ of 0.5.Isopropyl-β-D-thiogalactopyranoside (IPTG) was added to 1 mM andcultures were shaken overnight at 30° C. Bacterial pellets wereharvested and resuspended in {fraction (1/50)}th of the initial culturevolume with osmotic shock buffer (500 mM sucrose, 1 mM EDTA, 100 mMTris, pH 8.00), incubated for 30′ at 4° C., and centrifuged at 16,000×gfor 15′ at 4° C. Fab-containing supernatants were dialyzed against PBSand used in agglutination experiments without further purification.

Anti-Rh(D) Antibody Binding Assays

The binding of anti-Rh(D) Fab/phage or soluble Fab molecules to normalor partial Rh(D) antigens was assessed by indirect agglutination assaysas described (Siegel et al., 1994, Blood 83:2334; Siegel et al., 1997,J. Immunol. Meth. 206:73). Briefly, 100 μl aliquots of phage-displayedFabs or soluble Fabs were incubated with 50 μl of a 3% suspension ofRBCs. Following a one-hour incubation at 37° C., the RBCs were washed 3times with 2 ml of cold PBS to remove unbound antibody. The resultingRBC pellets were resuspended in 100 μl of a 10 μg/ml solution of sheepanti-M 13 antibody (5 Prime-3 Prime, Boulder, Col.) for Fab/phageexperiments or goat anti-human κ or λ light chain antibody (Tago,Burlingame, Calif.) for γ₁κ or γ₁λ soluble Fab experiments,respectively. The RBC suspensions were transferred to the round-bottomedwells of a 96-well microplate and left undisturbed for 2 hours. Negativereactions show sharp ˜2 millimeter diameter RBC spots whereas the RBCsin agglutinated wells form a thin carpet coating the entire floor of thewell (Siegel et al., 1997, J. Immunol. Meth. 206:73). Agglutinationtiters for recombinant antibodies were determined by performing serial2-fold dilutions in 1% BSA/PBS. Typically, Fab/phage had agglutinationtiters of 1/1024 to 1/2048 (where “neat” is defined as 5×10¹² cfu/ml;Siegel et al., 1997, J. Immunol. Meth. 206:73) and soluble Fabs hadagglutination titers of 1/64 to 1/128 when prepared as described above.

For determining Rh(D) epitope specificity for anti-Rh(D) Fab/phageantibodies, the following reference Rh(D) variant cells were used:O/D^(IIIa)Cce, G positive; B/D^(IIIc)Cce; A/D^(IVa)ce; A/D^(IVa)ce;O/D^(IVa)ce; O/D^(IVb)Cce; B/D^(IVb)Cce, Go^(a) negative, Rh32 negative;O/D^(Va)Cce; O/D^(Va)cEe, D^(w) positive; O/D^(VI)Cce; B/D^(VI)Cce;AB/D^(VI)Cce; A/D^(VI)cEe; O/D^(VII)Cce; and O/D^(VII)Cce. EachFab/phage antibody was tested on at least 3 separate occasions againstat least 2 different examples of each variant cell type and identicalepitope assignments were obtained each time. For antibodies thatdemonstrated not-previously-described patterns of reactivity orrepeatedly weak reactivity against one type of cell, monoclonalFab/phage were prepared on a least 4 separate occasions to verify thepatterns of reactivity.

For inhibition studies, the ability of antibodies with different Rh(D)epitope specificities to compete with each other for binding wasassessed by preparing stocks of each clone in both a soluble Fab formand a phage-displayed form. Pair-wise combinations of soluble Fabs andFab/phage were prepared and added to Rh(D)-positive RBCs. The resultingincubation mixes comprised 50 μl of a 3% suspension of RBCs, 100 μl ofundiluted soluble Fab, and 100 μl of Fab/phage diluted to its highestagglutinating titer. Following a 1-hour incubation at 37° C., RBCs werewashed, resuspended in anti-M13 antibody, and placed in microplate wellsas described above. That the amount of soluble Fab present in anincubation mixture was sufficient to compete away a Fab/phage thatshared the same binding site was determined by verifying that eachsoluble Fab preparation could block its own Fab/phage.

Inhibition experiments were also performed using pair-wise combinationsof soluble Fabs instead of soluble Fab and Fab/phage combinations. Inthis type of experiment, pairs of soluble Fabs specific for differentepitopes were chosen such that one Fab contained a λ light chain and theother a κ light chain. Incubations with RBCs were performed with one Fabin excess and the other in limiting amounts. Blocking of the latterantibody was assessed using a secondary antibody (anti-λ or anti-κ)specific for its light chain isotype.

Nucleotide Sequencing and Analysis

Plasmid DNA for sequencing was prepared using the Qiawell™ system(Qiagen, Chatsworth Calif.). Double-stranded DNA was sequenced usinglight chain or heavy chain immunoglobulin constant region reverseprimers or a set of unique pComb3H vector primers that anneal 5′ to therespective immunoglobulin chain (Barbas et al., 1991, Methods: ACompanion to Meth. Enzymol. 2:119; Roben et al., 1995, J. Immunol.154:6437) and automated fluorescence sequencing (Applied Biosystems,Foster City, Calif.). Sequence analysis and variable region germlineassignments were performed using DNAplot (Althaus et al., 1996, DNAPLOT,http://www.mrc_cpe.cam.ac.uk/imt_doc/DNAsearch.html) and the V BaseDirectory of Human V Gene Sequences (March 97 update; Tomlinson et al.,1996, V Base Directory of Human V Gene Sequences,http://www.mrc_cpe.cam.ac.uk/imt_doc/vbase_home_page.html). Germlineassignments were corroborated with the MacVector (v. 6.0) softwarepackage (Oxford Molecular Group, Oxford, UK) against the same database.Multiple sequence alignments and predictions of isoelectric point werecalculated using the Pileup and Isoelectric programs of the GCG softwarepackage (v. 8.0.1; GCG, Madison Wis.). Statistical analysis wasperformed with Statview (Abacus Concepts, Berkeley Calif.).

The results of the experiments presented in this Example are nowdescribed.

Sequence Analysis of Anti-Rh(D) Heavy and Light Chains

Example 1 describes the use of Fab/phage display and cell-surfacepanning to isolate a large array of anti-Rh(D) antibodies from theperipheral blood lymphocytes of a single hyperimmunized donor. Separateγ₁κ and γ₁λ Fab/phage display libraries were constructed and contained7×10⁷ and 3×10⁸ independent transformants, respectively, based onelectroporation efficiency. Each library was panned independently usinga simultaneous positive/negative selection strategy withmagnetically-labeled Rh(D)-positive RBCs and unmodified Rh(D)-negativeRBCs as described. Following two rounds of panning, 32 of 36 γ₁λ and 15of 15 γ₁κ clones were positive for anti-Rh(D) activity. After the thirdround of panning, 24 out of 24 γ₁λ and 12 out of 12 γ₁κ clones werepositive. Nucleotide sequencing of the 83 positive clones revealed atotal of 28 unique heavy and 41 unique light chains. Due tocombinatorial effects during phage display library construction, heavyand light chain gene segments paired to produce 53 unique Fabantibodies.

Anti-Rh(D) Heavy Chains

All of the heavy chain sequences used V_(H)III family-encoded geneproducts, as indicated in FIGS. 7 and 8. Several heavy chain sequencesshared identical VDJ joining regions, and 12 unique VDJ rearrangementswere identified. These rearrangements were designated VDJ1 throughVDJ12. Alignment of these sequences against the V Base Directory ofHuman V Gene Sequences revealed that only four V_(H)III genes were usedby these antibodies: VH3-21, VH3-30, VH3-33, and VH 3-30.3. VH3-21 wasused by 1 of the 12 VDJs and 2 of the 28 clones; VH3-30 was used by 1VDJ and 6 clones; VH3-33 was used by 9 VDJs and 19 clones; and VH3-30.3was used by 1 VDJ and 1 clone. Interestingly, VH3-30, VH3-33, andVH3-30.3 comprise a set of closely related genes (>98% homology; FIG.8B) and their next nearest neighbor, VH3-07, is only 90% homologous(FIG. 8C). Hereafter, these three genes are referred to as the “VH3-33superspecies”. Heavy chain E1 differed from VH3-21 by six mutations anddiffered from VH3-48 by ten mutations; hence, it was assigned to theformer germline gene. Because there were no common mutations among theVH3-33 clones, it is highly probable that the donor possessed the VH3-33germline gene. However, we could not formally rule out gene duplicationwith allelic variants of VH3-33 or the existence of variant alleles ofthe other germline genes in the donor. The isolation of clones sharingmultiple VDJ joining regions strongly suggests that cloning artifactscannot account for the V_(H) restrictions observed.

Neither J_(H) nor D segments showed restriction. At least 9 different Dsegments were used and J_(H) gene utilization comprised J_(H)6 (5 VDJsand 9 clones), J_(H)4 (4 VDJs and 10 clones), J_(H)3 (2 VDJs and 8clones) and J_(H)5 (1 VDJ and 1 clone). All four V_(H) genes wereChothia class 1-3 (Chothia et al., 1992, J. Mol. Biol. 227:799), and theCDR3s showed a narrow range of length from 15 to 19 residues.

Because rearranged heavy chain genes demonstrate extensive diversity,clones sharing identical VDJ rearrangements are generally considered tohave arisen from the same clone. Based upon nucleotide alignment withthe germline genes, the ontogeny tree in FIG. 9 was constructed for the12 VDJs and 28 clones. By using the most parsimonious mutation scheme(i.e. postulating the minimum number of mutations), putativeintermediate antibodies were derived for several of the VDJs and weredesignated Ca, Cb, Da, Db, and Dc (FIGS. 8A and 9). Compared with theisolated heavy chain clones, which had between 6 and 23 nucleotidedifferences from their germline counterparts, these putativeintermediates had between 3 and 12 mutations from germline. Based uponthe ontogeny tree, the number of independent mutations could betabulated among the clones. The most commonly mutated residues were 52aand 58 (7 independent mutations), followed by residues 30, 31 and 50 (6mutations), and residue 55 (5 mutations). In the VH3-33 superspecies,residues 52a and 58 in CDR2 are tyrosine residues and residue 52a wasmutated to phenylalanine in 6 of the 11 VDJs derived from VH3-33superspecies V_(H) genes. Mutations at residue 58 comprised glutamate(3), aspartate (2), histidine (1) and asparagine (1). The AGY serines atresidues 30, 31 and 55 were mutated to a number of different aminoacids, although the AGY serine at 82b was conserved in all clones. Thevaline at residue 50 in the VH3-33 superspecies also had a diverse setof mutations. This distribution of “hot spots” is similar to that seenwith non-productive rearrangements as previously reported by Dörner etal (1997, J. Immunol. 158:2779).

Anti-Rh(D) Light Chains

Seventeen of the 18 κ light chains were from the V_(κ)I family and theremaining light chain originated from a V_(κ)II family member germlinegene (FIG. 10). Only four V_(κ) germline genes were used (15 clones werederived from DPK9 alone), and the κ light chain clones had between 1 and49 mutations from their corresponding V_(κ) germline genes. All five ofthe known J_(κ) genes were used and were each joined to the DPK9 gene inone or more clones. Because the light chains showed considerably lessdiversity in their joining regions than the heavy chains, it wasdifficult to assign common clonal origins. However, an ontogeny tree wasconstructed by grouping common V and J gene segments along with commonmutations. Based upon this analysis, the 18κ chains comprised at least10 different recombination events.

λ light chains were restricted by their J_(λ) gene usage but showed norestriction in their use of V_(λ) genes (FIG. 11). The 23λ light chainsall used the J_(λ)2Vasicek gene but were derived from V_(λ)I (12clones), V_(λ)III (5), V_(λ)VII (3), V_(λ)II (2) and V_(λ)IV (1) familygenes. The number of mutations ranged from 2 to 41 from the nearestgermline V_(λ) gene. Based upon common joining regions and mutations,these 23 1 light chains were derived from at least 13 different B cells.

Assessment of the Diversity of the Non-Panned Libraries

In order to determine whether the apparent restriction in gene usage ofthe anti-Rh(D) antibodies could have been due to pre-selection factors(i.e. cloning artifacts), the diversity of the non-panned γ₁κ and γ₁λFab/phage libraries was assessed. By sequencing 39 randomly-pickedclones, we determined that there were no duplicate heavy or light chainsequences, and that there was significant heterogeneity in V gene familyrepresentation before selection (FIG. 12). In fact, the variable regiongene family distribution was not unlike that found by otherinvestigators for IgG-secreting lymphocytes in adult peripheral blood(Stollar, 1995, Ann. NY Acad. Sci. 764:547). Furthermore, of the 14V_(H)III-encoded negative clones, only one used a VH3-33 superspeciesgermline gene (VH3-30.3); the other 13 were encoded by VH3-07 (3), 3-09(2), 3-15 (2), 3-48 (2), 3-72 (2), 3-23 (1), and DP-58 (1). Therefore,the restriction of the 83 anti-Rh(D) clones to the VH3-33, 3-30, 3-30.3and 3-21 genes is significant and not a result of skewed representationof certain germline genes within the originally constructed γ₁κ and γ₁λFab/phage libraries.

Heavy and Light Chain Contribution to Rh(D) Epitope Specificity

Because of the conformational dependency of Rh(D) antigenicity, Rh(D)“epitopes” have been classically defined through the use of RBCsobtained from rare individuals whose cells appear to produce Rh(D)antigens “lacking” certain epitopes. Examining the pattern ofagglutination of a particular anti-Rh(D) monoclonal antibody with suchsets of partial Rh(D) RBCs enables one to categorize that antibody'sfine specificity.

Monoclonal Fab/phage preparations were prepared in triplicate for eachof the 53 anti-Rh(D) clones and tested against a panel of Rh(D) categorycells IIIa/c, IVa, IVb, Va, VI, and VII. This panel of cells candifferentiate between the Rh(D) epitope specificities as described byLomas et al. (1989, Vox Sang 57:261; designated epitopes epD1, epD2,epD3, epD4, epD5, and epD6/7). Agglutination experiments using theFab/phage clones demonstrated five different patterns of reactivity,including a new pattern which had not been described in the originalstudy by Lomas et al. or in the more recently-described (Scott, 1996,Transfus. Clin. Biol. 3:333; Stollar, 1995, Ann. N.Y. Acad. Sci.764:547) 9-, 30-, or 37-epitope systems (as indicated by the datadepicted in FIGS. 13 and 14). Although nearly all Fab/phage gaveunequivocal agglutination reactions, a few antibodies gave repeatedlyweak patterns of reactivity against one of the panel cells. For thesereactions, monoclonal Fab/phage were prepared on at least 4 separateoccasions to verify the patterns of reactivity.

The most commonly-recognized epitope was epD6/7, against which 13 of theclones described herein were directed. Interestingly, monoclonalanti-Rh(D) clones isolated using conventional tissue culture methods aremost often specific for epD6/7 (Mollison et al., 1993, In: BloodTransfusions in Clinical Medicine, 9th ed., Blackwell Scientific,Oxford, U.K.). EpD2, epD1, and epD3 were recognized by 10, 7, and 2clones, respectively. Six clones agglutinated cells of categoriesIIIa/c, IVa, and VII, but not of categories IVb, Va, and VI, and weredesignated anti-“epDX”. This pattern is identical to epD1, except thatthe IVa cell is agglutinated. Three clones gave intermediate reactionswith cell IVa, but otherwise showed patterns consistent with epDX orepD1. These clones were designated epDX¹ or epD1^(X) depending onwhether this reactivity against cell IVa was stronger or weaker,respectively (see FIG. 14). Similarly, reaction patterns for epD1 andepD2 differ by a positive reaction with the category Va cell; therefore,one clone was given epD2¹ specificity because it gave only moderatereactivity against that cell. Such variable reactions against one ormore partial Rh(D) cells have been observed for anti-Rh(D) monoclonalantibodies produced through conventional tissue culture methods (Tippettet al., 1996, Vox Sang. 70:123).

Because of the reassortment of heavy and light chain gene segments thatoccurs during the construction of a phage display library, a number ofclones were isolated that shared either a heavy (e.g. E1) or light (e.g.M3) chain sequence (FIG. 14). Some heavy chains were found to havepaired with both κ and λ light chains (e.g. C1, D20) and eachdemonstrated anti-Rh(D) specificity. Interestingly, some heavy chains(e.g. E1, D12) mapped to different epitopes depending upon the lightchains with which they were paired. In particular, the light chains oftwo such clones, E1/M2 and E1/M3, differed by only three amino acidresidues (FIG. 11) and these differences appear to confer specificityfor epD2 vs. epD3.

Inhibition Studies

To investigate the topological relationships among the Rh(D) epitopes,inhibition studies were performed. Gorick et al. (1988, Vox Sang.55:165) used pairs of non-labeled and ¹²⁵I-labeled anti-Rh(D) monoclonalantibodies to demonstrate that antibodies to at least three differentRh(D) epitopes (subsequently identified as epD1, D6 and D7; Lomas etal., 1989, Vox Sang. 57:261) could inhibit one another. Recombinantantibodies to five Rh(D) epitopes were used to confirm and extend thosefindings (FIG. 15). In one series of experiments, the ability to expresseach antibody in both a soluble Fab as well as phage-displayed form wasexploited to determine whether a soluble Fab directed against oneepitope would inhibit the agglutination induced by an Fab/phage directedagainst a different epitope. Reciprocal pairs of soluble Fab andFab/phage specific for epD1, epD2, epD3, epD6/7, and epDX were tested.All ten combinations showed mutual inhibition patterns (illustrated inFIG. 15A for an anti-epD3/anti-epD6/7 combination). To show that thisinhibition was not due to non-specific factors, a control with anirrelevant RBC-binding recombinant antibody (an anti-blood group Bantibody) was performed (FIG. 15B). That sufficient inhibitory amountsof soluble Fab was present were first verified by demonstrating thateach soluble Fab could inhibit its own Fab/phage (FIGS. 15A and 15B;samples on diagonal). Similar results were obtained using pairs ofsoluble Fabs which differed in their light chain isotype composition(FIG. 15C).

Isoelectric Point Analysis of Anti-Rh(D) Antibodies

The restriction in V_(H) germline gene usage to only four V_(H)IIIfamily members was intriguing in light of their ability to conferspecificity to a number of Rh(D) epitopes. V_(H) germline gene segmentsused to encode anti-Rh(D) antibodies are among the most cationicsegments available in the human V_(H) repertoire which may be used toaccount for the relatively high pI of polyclonal anti-Rh(D)-containingantisera (Boucher et al., 1997, Blood 89:3277; Abelson et al., 1959, J.Immunol. 83:49; Frame et al., 1969, Immunology 16:277). Although thecationic nature of the antibodies may be important for binding to Rh(D),a constitutive net positive charge may be necessary to permeate thehighly negative RBC ζ potential, thus permitting antibody to contactantigen (Mollison et al., 1993, In: Blood Transfusion in ClinicalMedicine, 9th ed., Blackwell Scientific, Oxford, U.K.). In either case,analysis of the predicted pI for the 28 heavy chains and 41 light chainsisolated here showed an interesting phenomenon for the heavy chains, ascompared with the light chains. Using the pI interval scale of Boucheret al. (1997, Blood 89:3277), the average pI of the four germline V_(H)segments used to encode the 28 heavy chains is high (9.87±0.15),significantly higher than that of 39 randomly-picked, non-Rh(D) bindingclones from the original non-panned libraries (9.24±0.80, P<10⁻⁵).Similar to the results of Boucher et al., the addition of D and J_(H)segments and the introduction of somatic mutation did not significantlychange the pI of the average anti-Rh(D) heavy chain (9.81±0.33, P<0.37).However, for the light chains, the average pI of their germlinecounterparts was not cationic, but the light chains became so throughthe addition of J_(L) segments and somatic mutation. Overall, for all18κ and 23λ light chains, paired t-test analyses before and aftersomatic mutation showed a significant increase in net positive chargewhen comparing gerrnline V_(L) (6.63±1.47) with expressed V_(L)(7.28±1.51, P<10⁻³) or germline V_(L)J_(L) (7.43±1.47) with expressedV_(L)J_(L) (8.55±1.35, P<10⁻⁷). There was no significant increase in asimilar analysis of 16 non-Rh(D) binding clones (P<0.59 and P<0.19,respectively). Examination of the light chain sequences listed in FIGS.10 and 11 revealed that this increase in pI resulted from mutations thatnot only introduced positively-charged residues, but also eliminatedsome negatively-charged residues. There were 31 such events, 29 (91%) ofwhich occurred in the light chain CDR regions.

Conventional and Phage-Displayed Anti-Rh(D) Monoclonal Antibodies

The phage-display derived anti-Rh(D) clones were compared with thoseproduced by conventional tissue culture techniques (EBV-transformationand cell fusion). Despite the relatively small number ofpreviously-published sequences for IgG anti-Rh(D) antibodies (N=21) andthe fact that they were derived from over 10 different donors, there wassurprisingly good correlation between the two groups, as indicated inTable 3. Both cohorts demonstrated a predominance of V_(H)III-familyencoded germline genes, particularly those of the VH3-33 superspecies.CDR3 regions had similar lengths ranging from 15-19 residues forFab/phage antibodies and 16-20 for conventional monoclonal antibodies,although one heterohybridoma was an outlier, having a CDR3 length of 28residues. κ light chains were biased towards V_(κ)1 family members and λlight chains demonstrated the preferential use of the J_(λ)2Vasicekgene. The only qualitative discrepancy was in V_(λ) family usage whereFab/phage clones demonstrated a slight preference for V_(λ)I vs.V_(λ)III family members for conventional monoclonal antibodies. However,in both cohorts, DPL16 was used more often than any other λ light chaingene.

TABLE 3 Comparison of IgG Fab/phage library-derived anti-Rh(D)monoclonal antibodies prepared as described herein with those previouslyproduced by conventional tissue culture methods Attribute PreviouslyPublished* Current Study Heavy Chains (by clone)† (by VDJ) VH3 familyderived 12/21 (57%) 28/28 (100%) 12/12 (100%) VH3-33 superspecies‡/VH310/12 (83%) 26/28 (93%) 11/12 (92%) VH3-33/VH3 9/12 (75%) 19/28 (68%)9/12 (75%) VH3-21/VH3 1/12 (8%) 2/28 (7%) 1/12(8%) VH4-34 derived 2/21(10%) 0/28 (0%) 0/12 (0%) JH6 usage 15/21 (71%) 9/28 (32%) 5/12 (42%)CDR3 length 16-20 (28§) 15-19 κ Light Chains Vκ1 family derived/total ρ8/12 (67%) 17/18 (94%) Jκ1 usage/total ρ 4/12 (33%) 6/18 (33%) Jκ2usage/total ρ 4/12 (33%) 6/18 (33%) λ Light Chains Vλ1 familyderived/total λ 2/8 (25%) 12/23 (52%) Vλ3 family derived/total λ 5/8(63%) 5/23 (22%) DPL16 derived/V13 family 3/5 (60%) 4/5 (80%) Jλ2Vasicekusage/total λ 6/8 (75%) 23/23 (100%) Notes for Table 3 *Compiled from atotal of 21 sequences of IgG anti-Rh(D) antibodies isolated frommultiple subjects originally published by Bye et., Hughes-Jones et al.,Chouchane et al., and Boucher et al. and available from Genbank. Onelight chain (Oak-3) was not available in Genbank and was not included inthe assessment. †For heavy chains, left column tabulates each cloneseparately; right column tabulates clones on the basis ofshared V-D-Jjoining regions ‡VH3-33 superspecies defined as the group of VH3 familygermline genes comprising VH3-33, VH3-30, and VH30.3. §CDR3 lengthoutlier

It has been suggested in the literature that the VH4-34 (VH4.21)germline gene, a gene used by many autoantibodies and cold agglutinins,may play an important role in the immune response to Rh(D) (Silbersteinet al., 1991, Blood 78:2377; Pascuel et al., 1991, J. Immunol. 146:4385;Silverman et al., 1988, J. Exp. Med. 168:2361; Thompson et al., 1991,Scand. J. Immunol. 34:509). However, these conclusions arose from theanalysis of IgM monoclonal antibodies and only 2 of the 21 publishedanti-Rh(D) IgG sequences used VH4-34 (Bye et al., 1992, J. Clin. Invest.90:2481). In a related series of experiments, aliquots of the γ₁κ andγ1λ libraries obtained after the second and third rounds of selectionwere pooled and then panned against the VH4-34 specific ratanti-idiotypic monoclonal antibody (9G4; Stevenson et al., 1989, Br. J.Haematol. 72:9). Although VH4-34 encoded antibodies were successfullyenriched, the Fab/phage were not specific for Rh(D) and displayedserological characteristics similar to those of cold agglutinins.

Rh(D) Epitopes and Significance of Antibody Sequences

Since the initial report by Argall et al. in 1953 (J. Lab. Clin. Med.41:895), it has been recognized that rare individuals who type asRh(D)-positive can produce allo-anti-Rh(D) antibodies in response toRh(D) immunization by transfusion or pregnancy. This phenomenon wasexplained by hypothesizing that the Rh(D) antigen is a “mosaicstructure” and that these individuals were producing alloantibodies toparts of the mosaic they lack. By systematically examining patterns ofreactivity between their cells and sera, RBCs expressing partial Rh(D)antigens were divided into categories, each presumed to have a differentabnormality in their Rh(D) antigen. Through the subsequent use of indexpanels of monoclonal anti-Rh(D) antibodies, a series of epitopes weredefined of which the number and combination varied from one Rh(D)category to another. As new monoclonal antibodies were produced, theirreactivity profiles against these partial Rh(D) RBCs became the standardmethod for determining Rh(D) antibody epitope specificity. Molecularanalyses of partial Rh(D) phenotypes have shown that the Rh(D) genes inthese individuals have either undergone intergenic recombination withthe highly homologous Rh(CE) gene, or, less commonly, have sustainedpoint mutation(s) (Cartron et al., 1996, Transfus. Clin. Biol. 3:497).

As noted earlier, to investigate the topological relationships amongRh(D) epitopes, Gorick et al. performed competition experiments withRh(D) monoclonal antibodies and observed varying degrees of inhibition(Gorick et al., 1988, Vox Sang. 55:165). These results, when combinedwith those of Lomas et al. (1989, Vox Sang. 57:261), suggested a modelfor Rh(D) in which epitopes are spatially distinct yet demonstrate acertain degree of overlap as illustrated in FIG. 16A. This modelexplained how antibodies to two different Rh(D) epitopes (in this caseepD2 and epD3) could inhibit each other's binding to wild type Rh(D),and how a change in the structure of Rh(D) in category VI RBCs (asteriskin FIG. 16A) would cause the loss of epD2. However, based upon thisconcept of Rh(D) epitopes as distinct domains, one would expect thatantibodies against different epitopes of Rh(D) would be structurally andgenetically distinct as well. Thus, it was surprising that theanti-Rh(D) clones described herein demonstrated such marked restrictionin gene usage. For example, only two superspecies of V_(H) genes wereused despite specificities for 4 of the original 6 Rh(D) epitopesdescribed by Lomas et al. (1989, Vox Sang. 57:261). Furthermore,multiple specificities could arise from a single heavy chain dependingupon the light chain with which it was paired (e.g. E1 with M2, M3, L3,or L4). In addition, other clones repeatedly demonstrated variable weakreactivity against certain Rh(D) category RBCs that would affect theepitope specificities to which they were assigned (e.g. C1 with O1, M1,or J5).

Several hypotheses could account for these findings. The most simplisticinterpretation is that the heavy chain does not directly interact withthe antigen, but rather is responsible for bringing the antibody inclose proximity with the antigen. The specific interactions between thelight chain and the antigen would then determine the epitope specificityfor that antibody. In this regard, the data presented herein areconsistent with the observations of Boucher et al. (1997, Blood 89:3277)on the relative cationic nature of anti-Rh(D) heavy chains. However,because it was determined during the studies described herein that lightchains become cationic during somatic mutation, the charge of the entireantibody may play a role in its ability to bind, resulting in theselection and expansion of particular B-cell clones.

A more compelling hypothesis is that Rh(D) epitopes do not differspatially but differ only in the number and arrangement of contactresidues presented, as illustrated in FIG. 16B. In other words, the“footprints” of most, if not all, anti-Rh(D) antibodies are essentiallyidentical to one another. The genetic events which produce partial Rh(D)molecules result in the loss of certain critical key points of contactnecessary for some antibodies to bind; alternatively, they result in theformation of new structures that interfere with the binding of otheranti-Rh(D) immunoglobulins. For example, the introduction of a “ledge”in Rh(D) category VI cells (asterisk in FIG. 16B) does not interferewith the binding of an anti-epD3 antibody, but does prevent the bindingof anti-epD2. Therefore, category VI RBCs are said to have epD3 but“lack” epD2.

This model is consistent with the inhibition experiments describedherein (e.g. FIG. 15) and with those of Gorick et al. (1988, Vox Sang.55:165) and offers an explanation for the marked restriction in heavychain gene usage. This model also reconciles a mechanism by which oneheavy chain (e.g. E1) can confer binding to multiple epitopes and whysome of the recombinant anti-Rh(D) antibodies described herein, as wellas some conventionally-produced monoclonal antibodies (e.g. Tippett etal., 1996, Vox Sang. 70:123), display variable reactivity againstcertain categories of partial Rh(D) RBCs. From the antigen'sperspective, this model explains how a single point mutation in Rh(D)can result in the loss of multiple Rh(D) epitopes (such as T283I incategory HMi RBCs) and how the residues associated with the expressionof some epitopes appear to be distributed among nearly all theextracellular loops of Rh(D). It also provides an understanding as tohow ≧37 “epitopes” can fit on the relatively smallextracellularly-exposed surface of the Rh(D) molecule.

This concept of “coincident” epitopes is best exemplified by comparingthe E1/M2 and E1/M3 clones described herein. The only difference betweenthe reactivity of E1/M2 and E1/M3 is the ability of the latter antibodyto agglutinate Rh(D) category VI cells, as depicted in FIG. 13. Hence,E1/M2 is classified as an anti-epD2 and E1/M3 as an anti-epD3 antibody.Light chains M2 and M3 differ by only 3 residues: D82A, G95aA, and W96V,as indicated in FIG. 11. Therefore, some combination of these threeresidues confers reactivity against category VI cells. In other words,epD2 and epD3, as seen by the E1/M2 and E1/M3 antibodies, differ by thebinding constraints imposed by at most three mutations. If the modeldepicted in FIG. 16A were correct and the epitopes were independent,these mutations would have to cause enough structural alteration in theantibody combining site so that a completely separate epitope on thesame antigen would be recognized. It would seem unlikely that these 3mutations could cause such a change, especially given the lack ofinternal homology domains in Rh(D). Therefore, it is concluded that itis far more plausible that the footprints of these 2 antibodies areessentially identical, and that one or more of these mutations (e.g. thetryptophan in CDR3 of M2) prevent(s) the interaction of E1/M2 withcategory VI RBCs. Since other clones demonstrate that the light chaincan confer specificity against epD1, epD2, or epD3 (with the E1 heavychain); epD1 or epDX (with C5); and epD1, epD2, and epD6/7 (with D12),we suggest that all 5 of these epitopes have similar antibody combiningsites.

Immunologic and Clinical Implications of Proposed Model

The model depicted in FIG. 16B leads to additional predictionsconcerning the Rh(D) immune response beyond simply clarifying what ismeant by an Rh(D) epitope. It is commonly stated in the transfusionmedicine literature that individuals whose RBCs express partial Rh(D)antigens are free to make antibodies to the Rh(D) epitopes they lack(Mollison et al., 1993, In: Blood Transfusion in Clinical Medicine, 9thed. Blackwell Scientific, Oxford, U.K.). Therefore, an individual whoproduces category VI RBCs should be able to make anti-epD2 but notanti-epD3. If these epitopes were truly independent, then the immunerepertoire of the anti-epD2 antibodies made by a category VI individualwould be similar to those produced by an Rh(D)-negative person. However,to the immune system, epD2 and epD3 are not independent.

It is herein postulated that somatic mutation of an anti-epD3 antibodycan change its fine specificity to that of epD2 (or vice-versa, see FIG.16C). Suppose that the preferred way of making an anti-epD2 antibody isthrough an anti-epD3 intermediate. To an Rh(D)-negative individual, thisprocess can take place unimpeded. However, in a category VI individual,this route would be unfavorable because an anti-epD3 antibody would beself-reactive. As a result, such an individual would have to makeanti-epD2 antibodies by alternative routes or by tolerating some degreeof auto-reactivity in the process. With respect to the latter point, itis of interest to note that a transient production of auto-anti-Rh(D)frequently precedes or accompanies the early production ofallo-anti-Rh(D) in individuals who express partial Rh(D) antigens (Chownet al., 1963, Vox Sang. 8:420; Macpherson et al., 1966, J. Clin. Pathol.45:748; Beard et al., 1971, Med. Genet. 8:317; Cook 1971, Br. J.Haematol. 20:369; Holland et al., Transfusion 13:363 (Abstract); Issit,1985, In: Applied Blood Group Serology, 3rd ed., Montgomery Scientific,Miami Fla.). It is predicted, therefore, that the anti-epD2 antibodiesfrom a category VI individual would be different in composition (i.e.gene usage) and quite possibly quantitatively depressed as compared toan Rh(D)-negative individual. This may be analogous to the antibodies ofthe ABO blood group system in which it has been observed that anti-A andanti-B titers in blood group O individuals are significantly higher thanin blood group B or A individuals, respectively (Ichikawa, 1959, Jap. J.Med. Sci. Biol. 12:1). Blood group O individuals are unconstrained increating their anti-A and anti-B immune repertoires while individualswho produce A or B antigens (2 nearly identical structures) must do soin a manner that avoids self-reactivity.

In the case of antibodies E1/M2 and E1/M3, they appear to have arisenfrom a common precursor B cell rather than directly from each other(FIG. 11). To test the framework of the hypothesis presented herein,i.e. somatic mutation resulting in “epitope migration” of an antibody,one may construct the precursors and potential intermediates between theM2 and M3 light chains and then determine what Rh(D) epitopespecificities (if any) they express. This concept of epitope migrationhas been previously reported for murine anti-cryptococcal and anti-typeII collagen antibodies (Mukherjee etal., 1995, J. Exp. Med. 181:405; Moet al., 1996, J. Immunol. 157:2440).

If the model proposed herein for Rh(D) epitopes is correct, then thequestion of the number of epitopes may be obsolete. There may be as manyepitopes as can be differentiated by the number of cell categories, i.e.2^(n) epitopes where n is the number of distinct partial Rh(D) RBCs.

A more important question is the interrelationships between the variousepitopes. For example, are some epitopes “further away” than others—notin the topological sense, but in terms of the number of mutational hitsan antibody needs to receive in order to change its serologicreactivity. Furthermore, does the humoral immune response in a partialRh(D) individual differ from that in an Rh(D)-negative individual in themanner predicted by this model? One may find that allo-anti-Rh(D)antibodies made by partial Rh(D) individuals are not as clinicallysignificant, i.e. capable of inducing hemolysis. This may explain whyhemolytic disease of the newborn due to anti-Rh(D) produced by pregnantindividuals with partial Rh(D) phenotypes is so rare even when takinginto account the low prevalence of the partial Rh(D) phenotypes(Mollison et al., 1993, In: Blood Transfusion in Clinical Medicine, 9thed. Blackwell Scientific, Oxford, U.K.). A better understanding of theimmune response to Rh(D) in these patients may alleviate concernsregarding the need to identify such individuals to ensure that they onlyreceive Rh(D)-negative blood products for transfusion and Rh(D)-immuneglobulin during pregnancy (Jones et al., 1995, Trans. Med. 5:171).Furthermore, with respect to the design of recombinant Rh(D)-immuneglobulin for use in Rh(D)-negative patients, it may not be necessary toformulate cocktails of monoclonal antibodies containing multiple Rh(D)epitope specificities.

Sequence Data

Genbank accession numbers for anti-Rh(D) heavy chains are as follows:

B01, AF044419; C01, AF044420; C03, AF044421; C04, AF044422; C05,AF044423; C08, AF044424; C10, AF044425; D01, AF044426; D03, AF044427;D04, AF044428; D05, AF044429; D07, AF044430; D08, AF044431; D09,AF044432; D10, AF044433; D11, AF044434; D12, AF044435; D13, AF044436;D14, AF044437; D15, AF044438; D16, AF044439; D17, AF044440; D18,AF044441; D20, AF044442; D30, AF044443; D31, AF044444; E01, AF044445;E03, AF044446.

Genbank accession numbers for anti-Rh(D) κ light chains are as

follows: F01, AF044447; G01, AF044448; H01, AF044449; I01, AF044450;I02, AF044451; I03, AF044452; I04, AF044453; I05, AF044454; I06,AF044455; I07, AF044456; I08, AF044457; I09, AF044458; I10, AF044459;I11, AF044460; I12, AF044461; I13, AF044462; I15, AF044463; I16,AF044464.

Genbank accession numbers for anti-Rh(D) λ light chains are as follows:J01, AF044465; J02, AF044466; J04, AF044467; J05, AF044468; K01,AF044469; K02, AF044470; K03, AF044471; L01, AF044472; L03, AF044473;L04, AF044474; L05, AF044475; M01, AF044476; M02, AF044477; M03,AF044478; N01, AF044479; N02, AF044480; O01, AF044481; O02, AF044482;O03, AF044483; P01, AF044484; Q01, AF044485; R01, AF044486; S01,AF044487.

Amino Acid Sequences of Anti-Rh(D) Heavy and Light Chains

The amino acid sequences of various anti-Rh(D) chains are representedusing single letter amino acid codes, as described herein.

The amino acid sequence of the anti-Rh(D) chain B01 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFRSYAMHWVRQAPGKGLEWVAATAYDGKNKYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVFYCARGGFYYDSSGYYGLRHYFDSWGQGTLVTVSS (SEQ ID NO: 1).

The amino acid sequence of the anti-Rh(D) chain C01 is

EVQLLESGGGVVQPGRSLRLSCAASGFSFSSYGMHWVRQAPGKGLEWVSVISYDGHHKNYADSVKGRFTISRDNSKKTLYLQMNSLRPEDTAVYYCANLRGEVTRRASVPFDIWGPGTMVTVSS (SEQ ID NO: 2).

The amino acid sequence of the anti-Rh(D) chain C03 is

EVQLLESGGGVVQHGRSLRLSCAASGFSFSSYGMHWVRQAPGKGLEWVSVISYDGHHKNYADSVKGRFTISRDNSKKTLYLQMNSLRPEDTAVYYCANLRGEVTRRASVPFDIWGPGTMVTVSS (SEQ ID NO: 3).

The amino acid sequence of the anti-Rh(D) chain C04 is

EVQLLESGGGVVQPGRSLRLSCAASGFSFSTYGMHWVRQAPGKGLEWVSVISYDGHNKNYADSVKGRFTISRDNSKKTLYLQMNSLRPEDTAVYYCANLRGEVTRRASIPFDIWGQGTMVTVSS (SEQ ID NO: 4).

The amino acid sequence of the anti-Rh(D) chain C05 is

EVQLLESGGGVVQPGRSLRLSCAASGFSFSSYGMHWVRQAPGKGLEWVAVISYDGTNKYFADSVKGRFTISRDNSKKTLYLQMTSLRPEDTAVYFCANLRGEVTRRASVPLDIWGQGTMVTVSS (SEQ ID NO: 5).

The amino acid sequence of the anti-Rh(D) chain C08 is

EVQLLESGGGVVQPGRSLRLSCAASGFSFSSYGMHWVRQAPGKGLEWVAVISYDGTNKYFADSVKGRFTISRDNSKKTLYLQMTSLRPEDTAVYFCANLRGEVTRRASVPLDIWGQGTMVTVSS (SEQ ID NO: 6).

The amino acid sequence of the anti-Rh(D) chain C10 is

EVQLLESGGGVVQPGRSLRLSCAASGFSFSSYGMHWVRQAPGKGLEWVSVISYDGHHKNYADSVKGRFTISRDNSKKTLYLQMNSLRPEDTAVYYCANLRGEVTRRASVPFDIWGPGTLVTVSS (SEQ ID NO: 7).

The amino acid sequence of the anti-Rh(D) chain D01 is

EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHWVRQAPGKGLEWVAVIWFDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRYLYYFDYWGQGTLVTVSS (SEQ ID NO: 8).

The amino acid sequence of the anti-Rh(D) chain D03 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWFDGSNKEYADSVKGRFTVSRDNSKNTLYLQMNSLRAEDTAVYYCAREEVVRGVILWSRKFDYWGQGTLVTVSS (SEQ ID NO: 9).

The amino acid sequence of the anti-Rh(D) chain D04 is

EVQLLESGGGVAQPGRSLRLSCVASGFSLRSYGMHWVRQAPGKGLEWVADIWFDGSNKDYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDWRVRAFSSGWLSAFDIWGQGTMVTVSS (SEQ ID NO: 10).

The amino acid sequence of the anti-Rh(D) chain D05 is

EVQLLEESGGGVAQPGRSLRLSCVASGFSLRSYGMHWVRQAPGKGLEWVADIWFDGSNKDYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDWRVRAFSSGWLSAFDIWGQGTTVSVSS (SEQ ID NO: 11).

The amino acid sequence of the anti-Rh(D) chain D07 is

EVQLLESGGGVVQPGRSLRLSCAVSGFTLTNYGMHWVRQAPGKGLEWVAHVWYDGSKTEYADSVKGRFAVSRDKSKNTLFLQMNSLTAEDTAIYYCARERREKVYILFYSWLDRWGQGTLVTVSS (SEQ ID NO: 12).

The amino acid sequence of the anti-Rh(D) chain D08 is

EVQLLEESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGRGLEWVALIWYDGGNKEYADSVKGRFSISRDNSKNTLYLQVNSLRADDTAVYYCARDQRAAAGIFYYSRMDVWGQGTTVTVSS (SEQ ID NO: 13).

The amino acid sequence of the anti-Rh(D) chain D09 is

EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGMHWVRQAPGKGLEWVAFIWFDGSNKYYEDSVKGRFTVSRDNSKNTLYLQMNSLRAEDTAVYYCAREGSKKVALSRYYYYMDVWGQGTTVTVSS(SEQ ID NO: 14).

The amino acid sequence of the anti-Rh(D) chain D10 is

EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGMHWVRQAPGKGLEWVAFIWFDGSNKYYEDSVKGRFTVSRDNSKNTLYLQMNSLRAEDTAVYYCAREVSKKVALSRYYYYMDVWGQGTTVTVSS (SEQ ID NO: 15).

The amino acid sequence of the anti-Rh(D) chain D11 is

EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGMHWVRQAPGEGLEWVAFIWFDGSNKYYADSVKGRFTVSRDNSKNTLYLQMNSLRAEDTAVYYCAREVSKKLALSRYYYYMDVWGQGTTVTVSS (SEQ ID NO: 16).

The amino acid sequence of the anti-Rh(D) chain D12 is

EVQLLESGGGVVQPGRSLRLACAASGFSFRSYGMHWVRQAPGRGLEWVAFTWFDGSNKYYVDSVKGRFTISRDNSKNTLYLEMNSLRVDDTAVYYCAREASMLRGISRYYYAMDVWGPGTTVTVSS (SEQ ID NO: 17).

The amino acid sequence of the anti-Rh(D) chain D13 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWFDGSNRDYAESVKGRFTISRDKSKNTLYLQMNSLRAEDSAVYYCARENVARGGGGVRYKYYFDYWGQGTLVTVSS (SEQ ID NO: 18).

The amino acid sequence of the anti-Rh(D) chain D14 is

EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWFDGSKRDYAESVKGRFTISRDNSKNTLYLQMNSLRAEDSAVYYCARENVARGGGGIRYKYYFDYWGQGTLVTVSS (SEQ ID NO: 19).

The amino acid sequence of the anti-Rh(D) chain D15 is

EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHWVRQAPGKGLEWVAVIWFDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRYLYYFDYWGQGTLVTVSS (SEQ ID NO: 20).

The amino acid sequence of the anti-Rh(D) chain D16 is

EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHWVRQAPGKGLEWVAVIWFDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRYLYYFDYWGQGTLVTVSS (SEQ ID NO: 21).

The amino acid sequence of the anti-Rh(D) chain D17 is

EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHWVRQAPGKGLEWVAVIWFDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRYLYYFDYWGQGTLVTVSS (SEQ ID NO: 22).

The amino acid sequence of the anti-Rh(D) chain D18 is

EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHWVRQASGKGLEWVAVIWFDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRYLYYFDYWGQGTLVTVSS (SEQ ID NO: 23).

The amino acid sequence of the anti-Rh(D) chain D20 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWFDGSNKEYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREEVVRGVILWSRKFDYWGQGTLVTVSS (SEQ ID NO: 24).

The amino acid sequence of the anti-Rh(D) chain D30 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMRWVRQAPGKGLEWVAVVYYDGSNKHYSDSVKGRFTISRDNSKNTLYLQMDSLRAEDTAVYYCARERNFRSGYSRYYYGMDVWGPGTTVTVSS (SEQ ID NO: 25).

The amino acid sequence of the anti-Rh(D) chain D31 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVVYYDGSNKHYSDSVKGRFTISRDNSKNTLYLQMDSLRAEDTAVYYCARERNFRSGYSRYYYGMDVWGPGTTVTVSS (SEQ ID NO: 26).

The amino acid sequence of the anti-Rh(D) chain E01 is

EVQLLESGGGLVKPGGSLRLSCAASGFTFSSYSMHWVRQAPGKGLEWVSSISNSNTYIYYADAVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARDSRYSNFLRWVRSDGMDVWGQGTTVIVSS (SEQ ID NO: 27).

The amino acid sequence of the anti-Rh(D) chain E03 is

EVQLLESGVESGGGLVKPGGSLRLSCAASGFTFSSYSMHWVRQGPGKGLEWVSSISNSNTYIYYADAVKGRFTISRDNAKNSLYLQMNSLRAEHTAVYYCARDSRYSNFLRWVRSDGMDVWGQGTTVIVSS (SEQ ID NO: 28).

The amino acid sequence of the anti-Rh(D) chain F01 is

AELTQSPSSLSASVGDRVTITCRASQGFRNDLGWYQQKPGKAPKRLIYATSSLQSGVPSRFSGSGSGTEFTLTINSLQPEDSATYYCLQHNSFPWTFGQGTKVEIKR (SEQ ID NO: 29).

The amino acid sequence of the anti-Rh(D) chain G01 is

AELTQSPLSLPVTPGEPASISCRSSQSLLHSSGFNFLDWYLQKPGQSPQLLIYMGSNRASGVPDRFSGSGSGTDFTLKINRVEAEDVGVYYCMQALQFPLTFGGGTKV EIKR (SEQ ID NO:30).

The amino acid sequence of the anti-Rh(D) chain H01 is

AELTQSPSFLSASVGDRVTITCRASQGITSYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTEFTLTIASLQPDDFATYYCQQLNNYPPFTFGPGTKVDIKR (SEQ ID NO: 31).

The amino acid sequence of the anti-Rh(D) chain I01 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPYTFGQGTKLEIKR (SEQ ID NO: 32).

The amino acid sequence of the anti-Rh(D) chain I02 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTLWTFGQGTKVEIKR (SEQ ID NO: 33).

The amino acid sequence of the anti-Rh(D) chain I03 is

AELTQSPSSLSASVADRVTITCRTSRNINRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTITSLQPEDFATYYCQQSYSTPFTFGPGTKVDLKR (SEQ ID NO: 34).

The amino acid sequence of the anti-Rh(D) chain I04 is

AELTQSPSSLSASVGDRVTITCRASQNIRRSLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSSNTPWTFGQGTKVEIKR (SEQ ID NO: 35).

The amino acid sequence of the anti-Rh(D) chain I05 is

AELTQSPSSLSASVGDRVTITCRASQSIRRYLNWYQHKPGKAPKLLIFAASSLQSGVPSRFTGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPQTFGQGTKVEIKR (SEQ ID NO: 36).

The amino acid sequence of the anti-Rh(D) chain I06 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPITFGQGTRLEIKR (SEQ ID NO: 37).

The amino acid sequence of the anti-Rh(D) chain I07 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPRTFGGGTKVEIKR (SEQ ID NO: 38).

The amino acid sequence of the anti-Rh(D) chain I08 is

AELTQSPFSLSASVGDRVTITCRASQTISRSLNWYQHKPGEAPKLLIYAASSLQRGVPPRFSGSGSGTDFTLTISSLQPEDFATYFCQQSVRIPYSFGQGTKLEIKR (SEQ ID NO: 39).

The amino acid sequence of the anti-Rh(D) chain I09 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDSTLTIS SLQPEDFATYYCQQLNSYPYTFGQGTKLEIKR (SEQ ID NO: 40).

The amino acid sequence of the anti-Rh(D) chain I10 is

AELTQSPSSLSASVGDRVTITCRASQNISSYLNWYQQKPGKAPKLLIYAASSLQSGVLSRFSGSGSGTDFTLTIS SLQPEDFATYYCQQSYSTPPYSFGQGTKLEIKR (SEQ ID NO: 41).

The amino acid sequence of the anti-Rh(D) chain I11 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPTLLINAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAIYYCQQRETFGQGTKLEIKR (SEQ ID NO: 42).

The amino acid sequence of the anti-Rh(D) chain I12 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPYTFGQGTKLEIKR (SEQ ID NO: 43).

The amino acid sequence of the anti-Rh(D) chain I13 is

AELTQSPSSLSASVGDRVTITCRASQSISRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYGTPHSFGRGTKLEIKR (SEQ ID NO: 44).

The amino acid sequence of the anti-Rh(D) chain I15 is

AELTQSPSSLSASVGDRVTITCRANQNIRRSLNWYQQKPGKAPNLLIYAASTLQGGVPSRFSGSGSGTDFTLTISSLQLADFATYYCQQTSATPWTFGQGTKVEIKR (SEQ ID NO: 45).

The amino acid sequence of the anti-Rh(D) chain I16 is

AELTQSPSSLPASVGDRVTITCRASQTIGFNLNWYQQTSGKPPKLLIYGVSKLQNGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQTNDALWTFGQGTKVEVRR (SEQ ID NO: 46).

The amino acid sequence of the anti-Rh(D) chain J01 is

AELQDPVVSVALGQTVRITCQGDGLRSYYASWYQQKPGQAPKLVMYGRNNRPSGIPGRFSGSSSGQTAALTITGTQAEDEADYYCQSRATSGNPVVFGGGTKLTV L (SEQ ID NO:47).

The amino acid sequence of the anti-Rh(D) chain J02 is

AELQDPVVSVALGQTVRITCQGDGLRSYYASWYQQKPGQAPKLVMYGRNNRPSGIPDRFSGSSSGQTAALTITGTQAEDEADYYCQSRATSGNPVVFGGGTKLTV L (SEQ ID NO:48).

The amino acid sequence of the anti-Rh(D) chain J04 is

AELQDPVVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNSRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRGSPHVAFGGGTKLTVL (SEQ ID NO: 49).

The amino acid sequence of the anti-Rh(D) chain J05 is

AELQDPVVSVALGQTVKITCQGDSLRKYYASWYQQKPGQAPVLVFYARNSRPSGIPDRFSGSNSGTTASLTIAGARAEDEADYYCHSRDSNGHHRVFGGGTKLTV L (SEQ ID NO: 50).

The amino acid sequence of the anti-Rh(D) chain K01 is

AELTQEPSLTVSPGGTVTLTCASSTGAVTSRYFPNWFQQKPGQAPRPLIYSASNKHSWTPARFSGSLLGGKAALTLSGVQPEDEAEYYCLLYYSGAWVFGGGTKLT VL(SEQ ID NO: 51).

The amino acid sequence of the anti-Rh(D) chain K02 is

AELTQEPSLTVSPGGTVTLTCASSTGAVTSRYFPNWFQQKPGQAPRPLIYSASNKHSWTPARFSGSLLGGKAALTLSGVQPEDEAEYYCLLYYSGAWVFGGGTKLT VL (SEQ ID NO: 52).

The amino acid sequence of the anti-Rh(D) chain K03 is

AELTQPPSLTVSPGGTVTLTCASSTGAVTSRYFPNWFQQKPGQAPRALIYGSNNKHSWTPARFSGSLLGGKAALTLSGVQPEDEAEYYCLLFYAGAWAFGGWTKLT VL (SEQ ID NO: 53).

The amino acid sequence of the anti-Rh(D) chain L01 is

AELTQPPSASGTPGQRVTISCSGGSSNIASNTVNWYQQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSATLVITGLQTGDEADYYCGTWDHSRSGAVFGGGTKLT VL (SEQ ID NO:54).

The amino acid sequence of the anti-Rh(D) chain L03 is

AELTQPPSASGTPGQRVTISCSGSSSNIGNNHVSWYQQLPGMAPKLLIYSNGQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWHDSLYGPVFGGGTKLT VL (SEQ ID NO:55).

The amino acid sequence of the anti-Rh(D) chain L04 is

AELTQPPSASGTPGQRVSISCSGSSSNIGSNTVNWYQQLPGTAPKLLISTNNQGPSGVPDRFSGSKSGTSSSLAISGLRSEAEDDYYCAAWDDTLNGVVFGGGTKLTV L (SEQ ID NO: 56).

The amino acid sequence of the anti-Rh(D) chain L05 is

AELTQPPSASGTPGLRVTISCSGSSSNIGSNIVNWYQQLPGTAPKLLIFSNNKRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCATWDDSLNGRVFGGGTKLTVL (SEQ ID NO: 57).

The amino acid sequence of the anti-Rh(D) chain M01 is

AELTQPPSASGTPGQRVTISCSGSNFNIGSNYVFWYQHVPGTAPKLLIYNNNQRPSGVPDRLSGSKSGASASLAINGLRSDDEADYYCTGWDDRLSGLIFGGGPKVT VL (SEQ ID NO:58).

The amino acid sequence of the anti-Rh(D) chain M02 is

AELTQPPSASGTPGQRVTISCSGSSSNIGSNYVYWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGWVFGGGTKLT VL (SEQ ID NO:59).

The amino acid sequence of the anti-Rh(D) chain M03 is

AELTQPPSASGTPGQRVTISCSGSSSNIGSNYVYWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEAEADYYCAAWDDSLSAVVFGGGTKLT VLL (SEQ ID NO:60).

The amino acid sequence of the anti-Rh(D) chain N01 is

AELTQPPSVSAAPGQKVTISCSGSSSNIDSNYVSWYQQLPGTAPKLLIFDNYRRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYYCATWDDSLNGRVFGGGTKLTV L (SEQ ID NO: 61).

The amino acid sequence of the anti-Rh(D) chain N02 is

AELTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLSAGRVRRMFGGG TKLTVLG (SEQ IDNO: 62).

The amino acid sequence of the anti-Rh(D) chain O01 is

AELTQPPSVSGAPGQRVTISCTGSSSNIGAPYGVHWYQQFPGTAPKLVIYNDNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGRVFGGGTKLT VL (SEQ ID NO:63).

The amino acid sequence of the anti-Rh(D) chain O02 is

AELTQPPSVSGAPGQTVTISCTGSSSSIGARYDVHWYQHLPGTAPKLLIYGNHNRPSGVPDRFSGSKSGTSASLAITGLQAEDEAEYYCQSYDNSLSGSSVFFGGGTK LTVL (SEQ ID NO:64).

The amino acid sequence of the anti-Rh(D) chain O03 is

AELTQPPSGAPGQTVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGPYVVFGGGTKLT VL (SEQ ID NO:65).

The amino acid sequence of the anti-Rh(D) chain P01 is

AELTQPPSVSVAPRQTARITCGGDKIGSNTVHWYRQMSGQAPVLVIYEDKKRPPGIPERFSGSTSGTTATLSISGAQVEDEADYYCYSRDNSGDQRRVFGAGTKLTV L (SEQ ID NO:66).

The amino acid sequence of the anti-Rh(D) chain Q01 is

AELTQPPSATASLGGSVKLTCILQSGHRNYAVAWHHQEAGKGPRFLMTVTNDGRHIKGDGIPDRFSGSASGAERYLSISGLQSEDEGDYYCQTWGTGMHVFGGGT KLTVL (SEQ ID NO:67).

The amino acid sequence of the anti-Rh(D) chain R01 is

AELTQPPSASGSPGQSVTISCTGASSDVGAYKHVSWYQQHPGKAPKLLTHEGTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSFAGNSVIFGGGTKLT VL (SEQ ID NO:68).

The amino acid sequence of the anti-Rh(D) chain S01 is

AELTQPPSVSGSPGQSITISCSDVGNYNLVSWYQQYPGKAPKLIIYEGSKRPSGVSSRFSGSRSGNTASLTISGLQAEDEADYHCCSYAISSRIFGGGTKLTVL (SEQ ID NO: 69).

Nucleotide Sequences of Anti-Rh(D) Heavy and Light Chains

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain B01 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGGAGCTATGCTATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGCTACAGCATATGATGGAAAAAATAAATACTACGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTTTCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTTTTACTGTGCGAGAGGCGGATTTTACTATGATAGTAGTGGTTATTACGGCTTGAGGCACTACTTTGACTCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA (SEQ ID NO: 70).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain C01 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGTCAGTTATATCATATGATGGACATCATAAAAACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAAAACGCTGTACCTGCAAATGAACAGCCTGAGACCTGAGGACACGGCTGTATATTACTGTGCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCTGTTCCCTTTGATATCTGGGGCCCAGGGACAAT GGTCACCGTCTCTTCA(SEQ ID NO: 71).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain C03 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGTGTGGTCCAGCATGGGAGGTCCCTGAGACTGTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGTCAGTTATATCATATGATGGACATCATAAAAACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAAAACGCTGTACCTGCAAATGAACAGCCTGAGACCTGAGGACACGGCTGTATATTACTGTGCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCTGTTCCCTTTGATATATGGGGCCCAGGGACAAT GGTCACCGTGTCTTCA(SEQ ID NO: 72).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain C04 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTACCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGTCAGTTATATCATATGATGGACATAATAAAAACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAAAACGCTGTACCTGCAAATGAACAGCCTGAGACCTGAGGACACGGCTGTGTATTACTGTGCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCTATTCCTTTTGATATCTGGGGCCAAGGGACAAT GGTCACCGTCTCTTCA(SEQ ID NO: 73).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain C05 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCAGCTTCAGTAGTTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCGTATGATGGAACTAATAAATACTTTGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAAAACGCTGTATCTGCAAATGACCAGCCTGAGACCTGAGGACACGGCTGTGTATTTCTGTGCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCCGTACCTCTTGATATCTGGGGCCAAGGGACAAT GGTCACCGTCTCTTCA(SEQ ID NO: 74).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain C08 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCAGCTTCAGTAGTTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCGTATGATGGAACTAATAAATACTTTGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAAAACGCTGTATCTGCAAATGACCAGCCTGAGACCTGAGGACACGGCTGTGTATTTCTGTGCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCTGTACCTCTTGATATCTGGGGCCAAGGGACAAT GGTCACCGTCTCTTCA(SEQ ID NO: 75).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain C10 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGTCAGTTATATCATATGATGGACATCATAAAAACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAAAACGCTGTACCTGCAAATGAACAGCCTGAGACCTGAGGACACGGCTGTATATTACTGTGCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCTGTTCCCTTTGATATCTGGGGCCCAGGGACATTG GTCACCGTCTCTTCA(SEQ ID NO: 76).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D01 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTGTCTGGTTTCACCTTCAATAACTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATTTGGTTTGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACACTGTACCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTATATTACTGTGCGAGAGAGAACCAGATAAAGCTATGGTCCCGATACCTTTACTACTTTGATTACTGGGGCCAGGGAAC CCTGGTCACCGTCTCCTCA(SEQ ID NO: 77).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D03 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTACCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGACTGGAGTGGGTGGCAGTTATATGGTTTGATGGAAGTAATAAGGAATATGCAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGAGACAATTCCAAGAACACGCTGTATCTACAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAGAAGTGGTTCGGGGAGTTATCTTATGGTCTCGGAAGTTTGACTACTGGGGCCAGGGAA CCCTGGTCACCGTCTCCTCA(SEQ ID NO: 78).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D04 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGCCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGCGTCTGGATTCAGCCTCAGGAGCTATGGCATGCACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGGAGTGGGTGGCAGATATATGGTTTGATGGAAGTAATAAAGATTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGTTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGATACGGCTGTGTATTATTGTGCGAGAGATTGGAGGGTGCGGGCCTTTAGTAGTGGCTGGTTAAGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCCTCA (SEQ ID NO: 79).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D05 is

GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCGTGGCCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGCGTCTGGATTCAGCCTCAGGAGCTATGGCATGCACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGGAGTGGGTGGCAGATATATGGTTTGATGGAAGTAATAAAGATTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGTTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTATTGTGCGAGAGATTGGAGGGTGCGGGCCTTTAGTAGTGGCTGGTTAAGTGCTTTTGATATCTGGGGCCAAGGGACCACGGTCAGCGTCTCCTCA (SEQ ID NO: 80).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D07 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGTGTCTGGATTCACCCTAACTAATTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCACATGTCTGGTATGATGGAAGTAAAACAGAATATGCAGACTCCGTCAAGGGCCGATTCGCCGTCTCCAGAGACAAATCCAAGAACACACTGTTTCTGCAAATGAACAGCCTGACAGCCGAGGACACGGCTATTTATTACTGTGCGAGAGAGAGGAGAGAGAAAGTCTATATATTGTTCTACTCGTGGCTCGACCGCTGGGGCCAGGGAA CCCTGGTCACCGTCTCCTCA(SEQ ID NO: 81).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D08 is

GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGGTTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAGGGGGCTGGAGTGGGTGGCTCTTATATGGTACGATGGAGGTAACAAAGAGTATGCAGACTCCGTGAAGGGCCGCTTCAGCATCTCCAGAGACAATTCCAAGAACACTCTGTATCTGCAAGTGAACAGCCTGAGAGCCGACGACACGGCTGTCTATTACTGTGCGAGAGACCAGAGAGCAGCAGCGGGTATCTTTTATTATTCCCGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 82).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D09 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGAAGCGTCTAAATTCACCCTCTACAATTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCATTTATATGGTTTGATGGAAGTAATAAATACTATGAAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAGGATCTAAGAAGGTGGCACTTTCTAGGTATTACTATTATATGGACGTCTGGGGCCAGGGGACCACGGTCACTGTCTCGTCA (SEQ ID NO: 83).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D10 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGAAGCGTCTAAATTCACCCTCTACAATTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCATTTATATGGTTTGATGGAAGTAATAAATACTATGAAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAGTATCTAAGAAGGTGGCACTTTCTAGGTATTACTACTATATGGACGTCTGGGGCCAGGGGACCACGGTCACTGTCTCCTCA (SEQ ID NO: 84).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D11 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGAAGCGTCTAAATTCACCCTCTACAATTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCGAAGGGCTGGAGTGGGTGGCATTTATATGGTTTGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAGTATCTAAGAAGCTGGCACTTTCTAGGTACTACTACTATATGGACGTCTGGGGCCAGGGGACCACGGTCACTGTCTCCTCA (SEQ ID NO: 85).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D12 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCGCCTGTGCAGCGTCTGGATTCAGCTTCAGGAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAGGGGGCTGGAGTGGGTGGCATTTACATGGTTTGATGGAAGCAATAAATATTATGTAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGGAAATGAACAGCCTGAGAGTCGATGACACGGCTGTATATTACTGTGCGAGAGAGGCGTCTATGCTTCGCGGAATTAGCAGATACTACTACGCGATGGACGTCTGGGGCCCAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 86).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D13 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTACTTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTTTGATGGAAGTAACAGAGACTATGCAGAGTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAAGTCCAAGAACACACTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACTCGGCTGTGTATTATTGTGCGAGAGAAAATGTGGCTCGTGGGGGGGGGGGCGTTCGATACAAGTACTACTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA (SEQ ID NO: 87).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D14 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTACTTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTTTGATGGAAGTAAGAGAGACTATGCAGAGTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAACTCCAAGAACACACTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACTCGGCTGTGTATTACTGTGCGAGAGAAAATGTGGCTCGTGGGGGGGGGGGCATTCGATACAAGTACTACTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA (SEQ ID NO: 88).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D15 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTGTCTGGATTCACCTTCAATAACTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATTTGGTTTGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACACTGTACCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTATATTACTGTGCGAGAGAGAACCAGATAAAGCTATGGTCCCGATACCTTTACTACTTTGACTACTGGGGCCAGGGAAC CCTGGTCACCGTCTCCTCA(SEQ ID NO: 89).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D16 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTGTCTGGTTTCACCTTCAATAACTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATTTGGTTTGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACACTGTACCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTATATTACTGTGCGAGAGAGAACCAGATAAAGCTATGGTCCCGATACCTTTACTACTTTGACTACTGGGGCCAGGGAAC CCTGGTCACCGTCTCCTCA(SEQ ID NO: 90).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D17 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTGTCTGGTTTCACCTTCAATAACTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATTTGGTTTGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACACTGTACCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTATATTACTGTGCGAGAGAGAACCAGATAAAGCTATGGTCCCGATACCTTTACTACTTTGACTACTGGGGCCAGGGAAC CCTGGTCACCGTCTCCTCC(SEQ ID NO: 91).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D18 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTGTCTGGTTTCACCTTCAATAACTATGGCATGCACTGGGTCCGCCAGGCTTCAGGCAAGGGGTTGGAGTGGGTGGCAGTTATTTGGTTTGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACACTGTACCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTATATTACTGTGCGAGAGAGAACCAGATAAAGCTATGGTCCCGATACCTTTACTACTTTGACTACTGGGGCCAGGGAAC CCTGGTCACCGTGTCCTCA(SEQ ID NO: 92).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D20 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTACCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGACTGGAGTGGGTGGCAGTTATATGGTTTGATGGAAGTAATAAGGAATATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTACAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAGAAGTGGTTCGGGGAGTTATCTTATGGTCTCGGAAGTTTGACTACTGGGGCCAGGGAA CCCTGGTCACCGTCTCCTCA(SEQ ID NO: 93).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D30 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCGCTGGGTCCGGCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTGTCTACTATGATGGAAGTAACAAACACTATTCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAACTCCAAGAACACGCTGTATCTACAAATGGACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAAGAAATTTTCGGAGTGGTTATTCCCGCTACTACTACGGTATGGACGTCTGGGGCCCAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 94).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain D31 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGGCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTGTCTACTATGATGGAAGTAACAAACACTATTCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAACTCCAAGAACACGCTGTATCTACAAATGGACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAAGAAATTTTCGGAGTGGTTATTCCCGCTACTACTACGGTATGGACGTCTGGGGCCCAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 95).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain E01 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCCTGGTCAAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGCTATAGCATGCACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTAATAGTAATACTTACATATACTACGCAGACGCAGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCACTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTACTACTGTGCGAGAGATTCTAGATACAGTAATTTCCTCCGTTGGGTTCGGAGCGACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCATCGTCTCCTCA (SEQ ID NO: 96).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain E03 is

GAGGTGCAGCTGCTCGAGTCTGGGGTGGAGTCTGGGGGAGGCCTGGTCAAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGCTATAGCATGCACTGGGTCCGCCAGGGTCCAGGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTAATAGTAATACTTACATATACTACGCAGACGCAGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCACTGTATCTGCAAATGAACAGCCTGAGAGCCGAGCACACGGCTGTGTACTACTGTGCGAGAGATTCTAGATACAGTAATTTCCTCCGTTGGGTTCGGAGCGACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCATCGTCTCCTCA (SEQ ID NO: 97).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain F01 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCTTTAGAAATGATTTAGGCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCGCCTGATCTATGCTACATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAACAGCCTGCAGCCTGAAGATTCTGCAACTTATTACTGTCTACAGCATAATAGTTTCCCGTGGACGTTCGGCCAAGGGACCAAG GTGGAAATCAAACGA (SEQID NO: 98).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain G01 is

GCCGAGCTCACTCAGTCTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCTAGTCAGAGCCTCCTGCATAGTAGTGGATTCAACTTTTTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGATCTATATGGGTTCTAATCGGGCCTCCGGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGCACAGATTTTACACTGAAAATCAACAGAGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAGCTCTACAATTTCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGA (SEQ ID NO: 99).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain H01 is

GCCGAGCTCACCCAGTCTCCATCCTTCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGGGCATTACGAGTTATTTAGCCTGGTATCAGCAAAAACCAGGGAAAGCCCCTAAGCTCCTAATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCGCCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGTCAACAGCTTAATAATTACCCCCCTTTCACTTTCGGCCCTGGGACCAA AGTGGATATCAAACGA(SEQ ID NO: 100).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I01 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTATCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCGTACACTTTTGGCCAGGGGACCA AGCTGGAGATCAAACGA(SEQ ID NO: 101).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I02 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCTGTGGACGTTCGGCCAAGGGACCAAGG TGGAAATCAAACGA (SEQID NO: 102).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I03 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGCGGACAGAGTCACCATCACTTGCCGGACAAGTCGGAACATTAACAGATACTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATTTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCACCAGTCTGCAACCTGAAGATTTTGCCACTTACTACTGTCAACAGAGTTACAGTACCCCTTTCACTTTCGGCCCTGGGACCAAAG TGGATCTCAAACGA (SEQID NO: 103).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I04 is

GCCGAGCTCACTCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAACATTAGGAGGTCTTTAAATTGGTATCAACAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGTTCCAATACCCCGTGGACGTTCGGCCAAGGGACCAAGG TGGAAATCAAACGA (SEQID NO: 104).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I05 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGGAGGTATTTAAATTGGTATCAGCACAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCACTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCAAACGTTCGGCCAAGGGACCAAGG TGGAAATCAAACGA (SEQID NO: 105).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I06 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCCGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCGATCACCTTCGGCCAAGGGACACGAC TGGAGATTAAACGA (SEQID NO: 106).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I07 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCGAACTTTCGGCGGAGGGACCAAGG TGGAGATCAAACGA (SEQID NO: 107).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I08 is

GCCGAGCTCACCCAGTCTCCATTCTCCCTGTCTGCATCTGTCGGAGACAGAGTCACCATAACTTGCCGGGCAAGTCAGACCATTAGCAGGTCTTTAAATTGGTATCAGCATAAACCAGGGGAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTCTGCAGCGTGGGGTCCCACCCAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGACTTTGCGACTTACTTCTGTCAACAGAGTGTCAGAATCCCGTACAGTTTTGGCCAGGGGACCAAGC TGGAGATCAAACGA (SEQID NO: 108).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I09 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTCCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTATTACTGTCAACAGCTTAATAGTTACCCGTACACTTTTGGCCAGGGGACCAAGC TGGAGATCAAACGA (SEQID NO: 109).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I10 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAACATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCTATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCGTATAGTTTTGGCCAGGGGACCA AGCTGGAGATCAAACGA(SEQ ID NO: 110).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I11 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTACGCTCCTGATCAATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATTAGCAGTCTGCAACCTGAAGATTTCGCAATTTACTACTGTCAACAGAGAGAAACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA CGA (SEQ ID NO: 111).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I12 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTATCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCGTACACTTTTGGCCAGGGGACCA AGCTGGAGATCAAACGA(SEQ ID NO: 112).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I13 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCCTCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACGGTACCCCTCACAGTTTTGGCCGGGGGACCAAGC TGGAGATCAAACGA (SEQID NO: 113).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I15 is

GCCGAGCTCACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAATCAGAACATTCGTAGATCTTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAACCTCCTGATCTATGCTGCATCCACATTGCAAGGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACTTGCGGATTTTGCAACTTACTACTGTCAACAGACTTCCGCTACCCCGTGGACGTTCGGCCAAGGGACCAAGG TGGAAATCAAACGA (SEQID NO: 114).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain I16 is

GCCGAGCTCACCCAGTCTCCATCGTCCCTGCCTGCATCTGTGGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGACTATTGGTTTTAATTTAAATTGGTATCAGCAAACATCTGGGAAGCCCCCTAAACTCCTAATCTATGGTGTTTCCAAGTTGCAAAATGGGGTCCCTTCACGGTTCAGTGGCAGTGGGTCCGGGACGGAATTCACCCTCACAATCAGCAGTCTGCAGCCTGAGGATTTTGCGACTTATTATTGTCAACAGACTAACGATGCGTTGTGGACGTTCGGCCAAGGGACCAAAGT GGAAGTCAGACGA (SEQID NO: 115).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain J01 is

GCCGAGCTCCAGGACCCTGTTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACTTGCCAAGGAGACGGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCGGGACAGGCCCCGAAACTTGTCATGTACGGTAGAAACAACCGGCCCTCAGGGATCCCAGGCCGATTCTCTGGCTCCAGCTCAGGGCAGACAGCTGCCTTGACCATCACGGGGACTCAGGCGGAGGATGAGGCTGACTATTACTGTCAGTCCCGTGCCACCAGCGGTAACCCTGTGGTGTTCGGCGGAGGGACTAA GCTGACCGTCCTG (SEQID NO: 116).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain J02 is

GCCGAGCTCCAGGACCCTGTTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACTTGCCAAGGAGACGGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCGGGACAGGCCCCGAAACTTGTCATGTACGGTAGAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGGCAGACAGCTGCCTTGACCATCACGGGGACTCAGGCGGAGGATGAGGCTGACTATTACTGTCAGTCCCGTGCCACCAGCGGTAACCCTGTGGTGTTCGGCGGAGGGACTAA GCTGACCGTCCTG (SEQID NO: 117).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain J04 is

GCCGAGCTCCAGGACCCTGTTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGCTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAGCCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCGTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCGGACTATTATTGTAGTTCGCGGGGCAGCCCCCACGTGGCATTCGGCGGAGGGACCAAACTGAC CGTCCTG (SEQ ID NO:118).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain J05 is

GCCGAGCTCCAGGACCCTGTTGTGTCTGTGGCCTTGGGACAGACAGTCAAGATCACATGCCAGGGAGACAGCCTCAGAAAGTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTGCTTGTCTTCTATGCTAGAAATAGCCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAACTCAGGAACCACAGCTTCCTTGACCATCGCTGGGGCTCGGGCGGAAGATGAGGCTGACTATTACTGTCACTCCCGGGACAGCAATGGTCACCATCGGGTGTTCGGCGGAGGGACCAA GCTGACCGTCCTA (SEQID NO: 119).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain K01 is

GCCGAGCTCACTCAGGAGCCCTCACTGACTGTGTCCCCAGGAGGGACAGTCACTCTCACCTGTGCTTCCAGCACTGGAGCAGTCACCAGTCGTTACTTTCCAAACTGGTTCCAGCAGAAACCTGGACAAGCACCCAGGCCACTGATTTATAGTGCAAGCAACAAACACTCCTGGACCCCTGCCCGGTTCTCAGGCTCCCTCCTTGGGGGCAAAGCTGCCCTGACACTGTCAGGTGTGCAGCCTGAGGACGAGGCTGAGTATTACTGCCTGCTCTACTATAGTGGTGCTTGGGTGTTCGGCGGAGGG ACCAAGTTGACCGTCCTT(SEQ ID NO: 120).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain K02 is

GCCGAGCTCACTCAGGAGCCCTCACTGACTGTGTCCCCAGGAGGGACAGTCACTCTCACCTGTGCTTCCAGCACTGGAGCAGTCACCAGTCGTTACTTTCCAAACTGGTTCCAGCAGAAACCTGGACAAGCACCCAGGCCACTGATTTATAGTGCAAGCAACAAACACTCCTGGACCCCTGCCCGGTTCTCAGGCTCCCTCCTTGGGGGCAAAGCTGCCCTGACACTGTCAGGTGTGCAGCCTGAGGACGAGGCTGAGTATTACTGCCTGCTCTACTATAGTGGTGCTTGGGTGTTCGGCGGAGGG ACCAAGCTGACCGTCCTA(SEQ ID NO: 121).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain K03 is

GCCGAGCTCACTCAGCCACCCTCACTGACTGTGTCCCCAGGAGGGACAGTCACTCTCACCTGTGCTTCCAGCACTGGAGCAGTCACCAGTCGTTACTTTCCAAACTGGTTCCAGCAGAAACCTGGCCAGGCACCCAGGGCACTGATTTATGGTTCAAACAACAAACACTCCTGGACCCCTGCCCGGTTCTCAGGCTCCCTCCTTGGGGGCAAAGCTGCCCTGACACTGTCAGGTGTGCAGCCTGAGGACGAGGCGGAGTATTACTGCCTGCTCTTCTATGCTGGTGCTTGGGCGTTCGGCGGATGGA CCAAGCTGACCGTCCTA(SEQ ID NO: 122).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain L01 is

GCCGAGCTCACGCAGCCGCCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGGAGGCAGCTCCAACATCGCAAGTAATACTGTAAACTGGTACCAGCAACTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGTAATAATCAGCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCACCCTGGTCATCACCGGGCTCCAGACTGGGGACGAGGCCGATTATTACTGCGGAACATGGGATCACAGCCGGAGTGGTGCGGTGTTCGGCGGAGGGACCAAACTGACCGTCTTA (SEQ ID NO: 123).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain L03 is

GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGGCAGTAGCTCCAACATCGGAAATAATCATGTAAGCTGGTACCAGCAACTCCCAGGAATGGCCCCCAAACTCCTCATCTATTCTAATGGTCAGCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGCGGCCTCCAGTCTGAGGATGAGGCTGATTATTATTGTGCAGCATGGCATGACAGCCTCTATGGTCCGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTC (SEQ ID NO: 124).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain L04 is

GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCAGCATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAATACTGTAAACTGGTACCAGCAGCTCCCAGGAACAGCCCCCAAACTCCTCATCTCTACTAATAATCAGGGGCCCTCAGGAGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCATCCTCCCTGGCCATCAGTGGGCTCCGGTCAGAGGCTGAGGATGATTATTACTGTGCAGCATGGGATGACACCCTGAATGGTGTGGTATTCGGCGGAGGGACCAAACTGACCGTCCTA (SEQ ID NO: 125).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain L05 is

GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGACTCCCGGGCTGAGGGTCACCATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAATATTGTAAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTTTAGTAATAATAAGCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTACTGTGCTACATGGGATGACAGCCTGAATGGTCGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA (SEQ ID NO: 126).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain M01 is

GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGCGGGTCACCATCTCTTGTTCTGGGAGCAACTTCAACATCGGAAGTAATTATGTATTCTGGTACCAGCATGTTCCAGGAACGGCCCCAAAACTCCTCATCTATAATAATAATCAACGCCCCTCTGGGGTCCCTGACCGACTCTCTGGCTCCAAGTCTGGCGCCTCAGCCTCCCTGGCCATCAATGGGCTCCGGTCCGATGATGAGGCTGATTATTACTGTACAGGATGGGATGACCGCCTGAGTGGCCTGATTTTCGGCGGAG GGCCAAAAGTGACCGTCCTA(SEQ ID NO: 127).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain M02 is

GCCGAGCTCACGCAGCCGCCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAATTATGTATATTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGCAGCATGGGATGACAGCCTGAGTGGTTGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA (SEQ ID NO: 128).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain M03 is

GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAATTATGTATACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATAGGAATAATCAGCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGCTGAGGCTGATTATTACTGTGCGGCATGGGATGACAGCCTGAGTGCCGTGGTATTCGGCGGAG GGACCAAACTGACCGTCCTA(SEQ ID NO: 129).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain N01 is

GCCGAGCTCACGCAGCCGCCCTCAGTGTCTGCGGCCCCAGGACAGAAGGTCACCATCTCCTGCTCTGGAAGCAGCTCCAACATTGACAGTAACTATGTATCCTGGTACCAGCAGCTCCCAGGAACAGCCCCCAAACTCCTCATTTTTGACAATTATAGGCGACCCTCAGGGATTCCTGACCGATTCTCAGGCTCCAAGTCTGGCACGTCAGCCACCCTGGGCATCACCGGACTCCAGACTGGGGACGAGGCCGATTATTACTGTGCAACATGGGATGACAGCCTGAATGGTCGGGTGTTCGGCGGAG GGACCAAGCTGACCGTCCTA(SEQ ID NO: 130).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain N02 is

GCCGAGCTCACGCAGCCGCCCTCAGTGTCTGCGGCCCCAGGACAGAAGGTCACCATCTCCTGCTCTGGAAGCAGCTCCAACATTGGGAATAATTATGTGTCCTGGTACCAGCAACTCCCAGGAACAGCCCCCAAACTCCTCATTTATGACAATAATAAGCGACCCTCAGGGATTCCTGACCGATTCTCTGGCTCCAAGTCTGGCACGTCAGCCACCCTGGGCATCACCGGACTCCAGACTGGGGACGAGGCCGATTATTACTGCGGAACATGGGATAGCAGCCTGAGTGCTGGCCGCGTTCGGCGGATGTTCGGCGGAGGGACCAAGTTGACCGTCCTGGGT (SEQ ID NO: 131).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain O01 is

GCCGAGCTCACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGAGGGTCACCATCTCCTGCACTGGGAGCAGCTCCAACATCGGGGCACCTTATGGTGTACACTGGTACCAGCAGTTTCCAGGAACAGCCCCCAAACTCGTCATCTACAATGACAACAATCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTGAGGATGAGGCTGATTATTACTGCCAGTCCTATGACAGCAGCCTGAGTGGAAGGGTGTTCGGCGGAGGGACCAAGCTGACCGTCCTA (SEQ ID NO: 132).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain O02 is

GCCGAGCTCACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGACGGTCACCATCTCCTGCACTGGGAGCAGCTCCAGCATCGGGGCACGTTATGATGTACACTGGTACCAACACCTTCCAGGAACAGCCCCCAAACTCCTCATCTATGGTAACCACAATCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTGAGGATGAGGCTGAATATTATTGCCAGTCCTATGACAACAGCCTGAGTGGTTCGTCTGTCTTTTTCGGCGGAGGGACCAAGCTGACCGTCCTA (SEQ ID NO: 133).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain O03 is

GCCGAGCTCACGCAGCCGCCCTCTGGGGCCCCAGGCCAGACGGTCACCATCTCCTGCACTGGGAGCAGCTCCAACATCGGGGCAGGTTATGATGTACACTGGTACCAGCAGCTTCCAGGAACAGCCCCCAAACTCCTCATCTATGGTAACAGCAATCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTGAGGATGAGGCTGATTATTACTGCCAGTCCTATGACAGCAGCCTGAGTGGTCCCTATGTGGTATTCGGCGGAGGGACCAAGCTGACCGTCCTA (SEQ ID NO: 134).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain P01 is

GCCGAGCTCACTCAGCCACCCTCGGTGTCAGTGGCCCCAAGACAGACGGCCAGGATTACCTGTGGGGGGGACAAAATCGGAAGTAACACTGTGCATTGGTACCGGCAGATGTCAGGCCAGGCCCCTGTTCTGGTCATCTATGAAGACAAAAAACGACCCCCCGGGATCCCTGAGAGATTCTCTGGTTCCACCTCAGGGACAACGGCCACCTTGAGTATCAGTGGGGCCCAGGTTGAGGATGAAGCTGACTACTACTGTTATTCAAGAGACAACAGTGGTGATCAGAGAAGGGTGTTCGGCGCAG GGACCAAGCTGACCGTCCTA(SEQ ID NO: 135).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain Q01 is

GCCGAGCTCACTCAGCCACCCTCCGCCACTGCCTCCCTGGGAGGCTCGGTCAAACTCACCTGCATTCTGCAGAGTGGCCACAGAAATTACGCCGTCGCTTGGCATCACCAAGAAGCAGGGAAGGGCCCGCGATTTTTGATGACGGTTACCAATGATGGCAGGCACATCAAGGGGGACGGGATCCCTGATCGCTTCTCAGGCTCCGCCTCTGGGGCTGAACGCTACCTCTCCATCTCCGGCCTCCAGTCTGAGGATGAGGGTGACTACTACTGTCAGACCTGGGGCACTGGCATGCATGTGTTCGGCGGAGGGACCAAACTGACCGTCCTA (SEQ ID NO: 136).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain R01 is

GCCGAGCTCACTCAGCCTCCCTCCGCGTCCGGGTCTCCTGGACAGTCAGTCACCATCTCCTGCACTGGAGCCAGCAGTGACGTTGGTGCTTATAAGCACGTCTCCTGGTACCAACAACACCCAGGCAAAGCCCCCAAACTCCTGACTCATGAGGGCACTAAGCGGCCCTCAGGGGTCCCTGATCGCTTCTCTGGCTCCAAGTCTGGCAACACGGCCTCCCTGACCGTCTCTGGGCTCCAGGCTGAGGATGAGGCTGATTATTACTGCAGCTCATTTGCAGGTAATTCCGTGATATTCGGCGGAGGG ACCAAGCTGACCGTCCTA(SEQ ID NO: 137).

The nucleotide sequence of the portion of the clone encoding theanti-Rh(D) chain S01 is

GCCGAGCTCACTCAGCCTCCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCAGTGATGTTGGGAATTATAACCTTGTCTCCTGGTACCAACAGTACCCAGGCAAGGCCCCCAAACTCATAATTTATGAGGGCAGTAAGCGGCCCTCAGGGGTTTCTAGTCGCTTCTCTGGCTCCAGGTCTGGCAACACGGCCTCCCTGACAATCTCTGGGCTCCAGGCTGAGGACGAGGCTGATTATCACTGCTGCTCATATGCAATTAGTAGCAGGATTTTCGGCGGAGGGACCAAGCTGACCG TCCTA (SEQ ID NO:138).

EXAMPLE 3 Isolation of Anti-Rh(D) Monoclonal Antibodies to Conventionaland Novel Epitopes Using a Heavy Chain/Light Chain Shuffling Approach

In view of the results obtained in Examples 1 and 2 herein, heavy andlight chains of antibodies of various Rh(D) epitope specificities wererandomly recombined in order to generate anti-Rh(D) antibodies havingadditional patterns of reactivity with Rh(D) variant cells. Using thisapproach, plasmid DNA obtained from the Fab/phage display librariesdescribed in panning rounds 2 and 3 of Example 1 was randomly recombinedto generate a “shuffled” Fab/phage display library. When the Rh(D)specificity of antibodies of this “shuffled” library was determine, itwas found that many of these antibodies exhibited novel epitopespecificity. Significantly, antibody clones having novel Rh(D) epitopespecificity were identified, including clones which bind to wild typeand certain partial D type red blood cells but which do not bind to Dcategory III red blood cells. The experiments described in this Exampletherefore demonstrate that the methods described in this specificationmay be used to generate antibody clones useful for diagnostic andtherapeutic applications in humans.

The materials and methods used in the experiments described in thisExample are now described.

Creation of Shuffled Fab/Phage Display Library

Two microgram aliquots of DNA obtained from libraries LP2, LP3, KP2, andKP3 (described herein in Example 1) were digested using the restrictionendonucleases SpeI and XhoI (15 and 60 units, respectively) in order todissociate DNA segments encoding individual (full length) heavy chainsfrom library plasmids encoding individual (full length) light chains.Endonuclease/DNA mixtures were incubated overnight at 37° C. After therestriction endonucleases were removed using standard phenol/chloroformand chloroform extraction techniques, the DNA was precipitated usingethanol.

Equivalent amounts of DNA from each of the four libraries (500 nanogramstotal) were mixed, and then the heavy chain-encoding DNA fragments werere-ligated into the library plasmids encoding individual light chains.This ligation was performed overnight at 20° C. in the presence of 3.5units of T4 DNA ligase in a total reaction volume of 70 microliters.This treatment generated re-ligated library plasmids encoding a lightchain and a heavy chain, wherein the light chain and the heavy chainwere not necessarily encoded by a single plasmid in the original libraryDNA. For this reason, the library of re-ligated plasmids was designateda “shuffled” library.

Three microliters of shuffled library suspension were mixed with analiquot of XL1-Blue electrocompetent cells (obtained from Stratagene, LaJolla, Calif.), and the cells were electroporated according to standardmethods. Electroporated cells were cultured on plates containing Luriabroth comprising 100 micrograms per milliliter carbenicillin.

Anti-Rh(D) Specificity of “Shuffled” Library Antibodies

Fifty-six randomly chosen colonies were selected, and monoclonalFab/phage preparations were separately produced from each of theseindividual colonies, using the methods described herein in Example 1.Rh(D) specificity was determined by indirect agglutination usinganti-M13 antibody, as described herein in Examples 1 and 2. Plasmid DNAwas separately prepared from each of the Fab/phage preparations whichexhibited Rh(D) specificity, and the DNA sequences encoding the heavyand light chains expressed by each preparation were determined asdescribed herein.

The results of the experiments presented in this Example are nowdescribed.

Anti-Rh(D) Specificity of “Shuffled” Library Antibodies

Of the 56 randomly-chosen “shuffled” library clones, 34 (61%)demonstrated specificity for Rh(D). The Rh(D) epitope specificity, theagglutination pattern, and the heavy and light chain sequences of these34 clones are listed in Table 4. Of these 34 clones, 19 exhibitedspecificity for previously-described Rh(D) epitopes (e.g. epD 1, epD 2,epD 6/7, and epD X), and one bound too weakly to wild-typeRh(D)-positive red blood cells to characterize is epitope specificity(i.e. clone SH44). However, 14 of the clones identified in Table 4exhibited novel Rh(D) epitope specificity. Some of these 14 antibodyclones comprised a heavy chain, a light chain, or both, that wereidentified herein in Examples 1 or 2. However, half (17/34) of the heavychain sequences and about 80% (28/34) of the light chain sequences hadnot been identified in Examples 1 or 2.

The Rh(D)-specific antibody clones isolated from the “shuffled” libraryare useful for characterizing and classifying patient red blood cellsthat express variant forms of the Rh(D) antigen. Of particular interestare clones SH18, SH20, and SH46. These three clones agglutinate wildtype red blood cells and certain partial D-type red blood cells, but donot agglutinate D category III red blood cells (a.k.a. partial Rh(D)IIIcells). It is believed that all previously-characterized humanmonoclonal anti-Rh(D) antibodies agglutinate D category III red bloodcells. Therefore these three clones are particularly useful fordifferentiating D category III red blood cells from other types of redblood cells.

From a clinical perspective, it has heretofore only been possible toretrospectively identify D category III red blood cells in a patientafter they have been erroneously presumed to have wild-typeRh(D)-positive cells. For example, transfusion of an individual having Dcategory III red blood cells with wild-type Rh(D) cells inducesproduction of anti-Rh(D) alloantibodies in the individual. Previously,the presence of D category III red blood cells in patients could only bedetermined by the production of such anti-Rh(D) alloantibodies in atransfusion recipient who does not naturally harbor D category III redblood cells. Although providing transfused blood comprising D categoryIII red blood cells to a patient who does not naturally harbor suchcells will not necessarily cause immediate harm to the patient, thepatient thereby becomes alloimmunized against D category III red bloodcells. Such alloimmunized individuals may develop complicationsincluding hemolytic transfusion reactions or hemolytic disease of thenewborn.

TABLE 4 Analysis of Anti-RH(D) Clones Obtained by Chain Shuffling. HEAVYCHAIN LIGHT CHAIN AGGLUTINATION PATTERN‡ Rh(D) CLONE SEQUENCE† SEQUENCE†wt III IVa IVb V V1 VII SPECIFICITY SH04 SEQ ID NOs: 24/93 SEQ ID NOs:35/104 + + + + + 0 + epD 6/7 SH08 SEQ ID NOs: 12/81 SBQ ID NOs:154/197 + + + + + 0 + epD 6/7 SH10 SEQ ID NOs: 139/182 SEQ ID NOs:47/116 + 0 0 0 0 0 0 novel SH12 SEQ ID NOs: 9/78 SEQ ID NOs:155/198 + + + + + 0 + epD 6/7 SH13 SEQ ID NOs: 26/95 SEQ ID NOs:156/199 + 0 0 0 0 0 0 novel SH14 SEQ ID NOs: 24/93 SEQ ID NOs:157/200 + + + + + 0 + epD 6/7 SH16 SEQ ID NOs: 140/183 SEQ ID NOs:158/201 + 0 + + 0 0 0 novel SH17 SEQ ID NOs: 141/184 SEQ ID NOs:47/116 + + 0 0 0 0 + epD 1 SH18 SEQ ID NOs: 142/185 SEQ ID NOs:159/202 + 0 + + 0 0 0 novel SH20 SEQ ID NOs: 143/186 SEQ ID NOs:160/203 + 0 + + + 0 0 novel SH21 SEQ ID NOs: 9/78 SEQ ID NOs:161/204 + + + 0 + 0 0 novel SH24 SEQ ID NOs: 144/187 SEQ ID NOs:162/205 + 0 0 0 0 0 0 novel SH25 SEQ ID NOs: 145/188 SEQ ID NOs:35/104 + + 0 0 + 0 + epD 2 SH26 SEQ ID NOs: 21/90 SEQ ID NOs:163/206 + + + 0 0 0 0 novel SH28 SEQ ID NOs: 146/189 SEQ ID NOs:164/207 + + 0 0 + 0 + epD 2 SH30 SEQ ID NOs: 12/81 SEQ ID NOs:165/208 + + + + + 0 + epD 6/7 SH32 SEQ ID NOs: 147/190 SEQ ID NOs:166/209 + 0 0 0 0 0 0 novel SH34 SEQ ID NOs: 5/74 SEQ ID NOs:167/210 + + 0 0 0 0 + epD 1 SH36 SEQ ID NOs: 14/83 SEQ ID NOs: 168/211 +0 0 0 0 0 0 novel SH37 SEQ ID NOs: 148/191 SEQ ID NOs: 50/119 + + + 0 00 + epD X § SH39 SEQ ID NOs: 149/192 SEQ ID NOs: 169/212 + 0 0 0 0 0 0novel SH41 SEQ ID NOs: 24/93 SEQ ID NOs: 170/213 + + + + + 0 + epD6/7SH44 SEQ ID NOs: 150/193 SEQ ID NOs: 171/214 + not determined SH46 SEQID NOs: 13/82 SEQ ID NOs: 172/215 + 0 + + 0 0 0 novel SH47 SEQ ID NOs:151/194 SEQ ID NOs: 173/216 + + 0 0 + 0 + epD 2 SH48 SEQ ID NOs: 6/75SEQ ID NOs: 174/217 + + 0 0 0 0 + epD 1 SH49 SEQ ID NOs: 17/86 SEQ IDNOs: 175/218 + + 0 0 0 0 + epD 1 SH50 SEQ ID NOs: 146/189 SEQ ID NOs:176/219 + + 0 0 0 0 + epD 1 SH51 SEQ ID NOs: 17/86 SEQ ID NOs:177/220 + + 0 0 + 0 + epD 2 SH52 SEQ ID NOs: 24/93 SEQ ID NOs:178/221 + + 0 0 0 0 + epD 1 SH53 SEQ ID NOs: 146/189 SEQ ID NOs:47/116 + 0 0 0 0 0 0 novel SH54 SEQ ID NOs: 152/195 SEQ ID NOs:179/222 + + 0 0 0 0 + epD 1 SH55 SEQ ID NOs: 21/90 SEQ ID NOs:180/223 + + 0 0 + 0 + epD 2 SH56 SEQ ID NOs: 153/196 SEQ ID NOs:181/224 + + 0 0 0 0 0 novel Notes for Table 4 †“SEQ ID NOs: A/B” meansthat the chain had amino acid sequence “A” and was encoded by nucleotidesequence “B”. ‡ “+” means agglutination occurred; “0” meansagglutination did not occur. *weak § as discussed in Example 2.

Amino Acid Sequences of Anti-Rh(D) Heavy and Light Chains

The amino acid sequences of various anti-Rh(D) antibody chains were asfollows, and are represented using single letter amino acid codes.

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH10 is

EVQLLEESGGGVVQPGRSLRLSCAASGFTFSRNGMHWVRQAPGKGLEWVAFIWFDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRADDTAVYYCAREEALFRGLTRWSYGMDVWGQGTTVSVSS (SEQ ID NO: 139).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH16 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGRGLEWVALIWYDGGNKEYADSVKGRFSISRDNSKNTLYLQVNSLRADDTAVYYCARDQRAAAGIFYYSRMDVWGQGTTVTVSS (SEQ ID NO: 140).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH17 is

EVQLLESGGGLVQPGGSLRLSCGASGIPFVSSWMAWVRQAPGKGLEWVANIKQDGSKKNYVDSVEGRFTISRDNAKNSLYLQMDSLRAEDTRIYYCARDSLTCFD YWGQGALVTVSS (SEQID NO: 141).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH18 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFRSYAMHWVRQAPGKGLEWVAATAYDGKNKYYADSVKGRFTISRDNSMNTLFLQMNSLRAEDTAVFYCARGGFYYDSSGYYGLRHYFDSWGQGTLVTVSS (SEQ ID NO: 142).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH20 is

EVQLLEESGGGVVQPGRSLRLSCAASGFTFRSYAMHWVRQAPGKGLEWVAVISYDGSTIYYADSVKGRFTISRANSKNTLFLQMNSLRTEDTAVYYCTRGGFYYDSSGYYGLRHYFDYWGQGTLVTVSS (SEQ ID NO: 143).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH24 is

EVQLLESGGGVAQPGRSLRLSCVASGFSLRSYGMHWVRQAPGKGLEWVADIWFDGSNKDYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDWRVRAFSSGWLSAFDIWGQGTMVTVSS (SEQ ID NO: 144).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH25

EVQLLEESGGGVVQPGRSLRLACAASGFSFRSYGMHWVRQAPGRGLEWVAFTWFDGSNKYYVDSVKGRFTISRDNSKNTLYLEMNSLRVDDTAVYYCAREAPMLRGISRYYYAMDVWGPGTTVTVSS (SEQ ID NO: 145).

The amino acid sequence of the heavy chain of each of anti-Rh(D)antibody clones SH28, SH50, and SH53 is

EVQLLESGGGGVQPGRSLRLSCAASGFTFNSYAMYWVRQPPGKGLEWVAAIWYDGSNKEYADFVKGRFTISRDNSKNTLSLQMNSLRDEDTAVYYCAREANLLRGWSRYYYGMDVWGQGTTVTVSS (SEQ ID NO: 146).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH32 is

EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGMHWVRQAPGKGLEWVAFIWFDGSNKYYEDSVKGRFTVSRDNSKNTLYLQMNSLRAEDTAVYYCARELSKKVALSRYYYYMDVWGQGTTVTVSS (SEQ ID NO: 147).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH37 is

EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGMHWVRQAPGKGLEWVAFIWFDGSNKYYEDSVKGRFTVSRDNSKNTLYLQMNSLRAEDTAVYYCARELSKKVALSRYYYYMDVWGQGTTVTVSS (SEQ ID NO: 148).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH39 is

EVQLLEQSGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWFDGSNKEYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREEVVRGVILWSRKFDYWGQGTLVTVSS (SEQ ID NO: 149).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH44 is

EVQLLESGGGVAQPGRSLRLSCVASGFSLRSYGMHWVRQAPGKGLEWVADIWFDGSNKDYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDWRVRAFSSGWLSAFDIWGQGTMVTVSS (SEQ ID NO: 150).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH47 is

EVQLLESGGGVVQPGRSLRLSCAASGFSFSNYAMHWVRQAPGKGLEWVAVTSFDGSIKDYADSVKGRFTISRDNSKNTLYLQMNSLRDEDTAVYYCARERGMIVVVRRRNAFDIWGQGTMVTVSS (SEQ ID NO: 151).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH54 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFSRNGMHWVRQAPGKGLEWVAFIWFDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRADDTAVYYCAREEALFRGLTRWSYGMDVWGQGTTVSVSS (SEQ ID NO: 152).

The amino acid sequence of the heavy chain of anti-Rh(D) antibody cloneSH56 is

EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVVYYDGSNKHYSDSVKGRFTIFRDNSKNTLYLQMDSLRAEDTAVYYCARERNFRSGYSRYYYGMDVWGPGTTVTVSS (SEQ ID NO: 153).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH8 is

AELTQSPSSLAASVGDRVTITCRANQTIRTSLNWYQQRPGKAPNLLIYGASRLHSGVPSRFSGGISGADFTLTISSLQPEDFATYYCQQTYGYSRTFGQGTKVDIKR (SEQ ID NO: 154).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH12 is

AELTQSPFSLSASVGDRVTITCRASHNIYRSLNWFQHKPGEAPKLLVYAASSLQRGVPTRFSGSGSGTDFTLTISSLQPEDSATYFCQQSVTFPYTFGQGTKLEIRR (SEQ ID NO: 155).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH13 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLRSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIKR (SEQ ID NO: 156).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH14 is

AELTQSPSSLSASVGDRVTITCRASQNIRRSLNWYQHKPGRAPRLLIYAASTLQSGVPSRFRGSGSGTDFTLTINSLQPADFATYYCQQSSNTPWTFGHGTKVEIKR (SEQ ID NO: 157).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH16 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGGGTKVEIKR (SEQ ID NO: 158).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH18 is

AELTQSPSSLSASVGDRVTITCRASQSISIALNWYQQRPGKAPKLLMYATSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYNKPTFGPGTKVDIKR (SEQ ID NO: 159).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH20 is

AELTQSPFSLSASVGDRVTITCRASQSISRSLNWYQHKPGEAPKLLIYAASSLQRGVPPRFSGSGSGTDFTLTISSLQPEDFATYFCQQSVRIPYSFGQGTKLEIKR (SEQ ID NO: 160).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH21 is

AELTQSPSFLSASVGDRVTITCRASQGIRSYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTEFTLTIASLQPDDFATYYCQQLNNYPPFTFGPGTKVDIKR (SEQ ID NO: 161).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH24 is

AELTQSPSSLSASVGDRVTITCRASQSISTYLNWYQQRPGKAPNLLIYAASTLQRGVPSRFTGSGSGTDFTLTISSLQPEDFATYYCQQSYTTLWTFGQGTKMEIRR (SEQ ID NO: 162).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH26 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSFRRYSFGQGTKLEIKR (SEQIDNO: 163).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH28 is

AELTQSPSSLSASVGDRVTITCRADQNIRRSLNWFQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSSSTPWTFGRGTKVEIKR (SEQ ID NO: 164).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH30 is

AELTQSPSSLSASVGDRVTITCRASQSIRRSLNWYQQSPGKTPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTLTFGGGTKVEIKR (SEQ ID NO: 165).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH32 is

AELTQEPSLTVSPGGTVTLTCASSTGAVTSRYFPNWFQQKPGQAPRALIYGSNNKHSWTPARFSGSLLGGKAALTLSGVQPEDEAEYYCLLFYAGAWAFGGGTKLT VL (SEQ ID NO:166).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH34 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASGLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPYTFGQGTKLEIKR (SEQ ID NO: 167).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH36 is

AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKSPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPAFGPGTKVDIKR (SEQ ID NO: 168).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH39 is

AELTQSPSSLSASVGDRVTITCRASQTIGRYLNWYQQRPGKAPKLLVYAVSSLQSGAPSRFSGSGSGTHFTLTITSLQPEDFATYFCQQSYSSPFTFGQGTKVEIKR (SEQ ID NO: 169).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH41 is

AELTQSPSSLSASVGDRVTITCRASQNIRRSLNWYQHKPGRAPRLLIYAASTLQSGVPSRFRGSGSGTDFTLTINSLQPADFATYYCQQSSNTPWTFGHGTKVEIKR (SEQ ID NO: 170).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH44 is

AELTQSPSSLSASVGDRVIITCRASQTIPRFLNWYQQKPGKAPVLLIHSISSLQSGVPSRFSASGSGTEFTLTISSLQPEDFATYYCQQSYSNLSFGPGTTVDIRR (SEQ ID NO: 171).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH46 is

AELTQSPSSLSASVGDRVTITCRASQYISSYLNWYQQKPGKAPNLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYSSPSTFGPGTKVDIKR (SEQ ID NO: 172).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH47 is

AELTQSPSSLSASVGDRVTITCRASQSISNYLNWYQQKPGKAPNLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSYPRTFGQGTKVEIRR (SEQ ID NO: 173).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH48 is

AELTQSPSSLSASVGDRVTITCRASQYISSYLNWYQQKPGKAPNLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTYSSPSTFGPGTKVDIKR (SEQ ID NO: 174).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH49 is

AELTQSPSSLSASVGDRVTVTCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPWTFGQGTKVEIKR (SEQ ID NO: 175).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH50 is

AELTQSPSSLSASVGDRVTVTCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPWTFGQGTKVEIKR (SEQ ID NO: 176).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH51 is

AELTQSPSFLSASVGDRVTITCRASQGIRSYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQLNNYPPFTFGPGTKVDIKR (SEQ ID NO: 177).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH52 is

AELTQSPGTLSLSPGERATLSCRASQSISSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIKR (SEQ ID NO: 178).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH54 is

AELTQSPSSMSASVGDRVTITCRASQSIGTYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPWTFGQGTKVEIKR (SEQ ID NO: 179).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH55 is

AELTQPPSASGTPGQRVTISCSGSSSNIGSKYVYWYQQLPGTAPKLLIYSNNQRPSGVPDRFSAFKSGTSASLAITGLQAEDEANYYCQSYDSGLSGWVFGGGTKLT VL (SEQ ID NO:180).

The amino acid sequence of the light chain of anti-Rh(D) antibody cloneSH56 is

AELTQSPSSLSASVGDRVTITCRASQSISRYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFALTISSLLPEDFATYYCQQGYSTPPYSFGQGTKLEIKR (SEQ ID NO: 181).

Nucleotide Sequences of Anti-Rh(D) Heavy and Light Chains

The nucleotide sequences encoding various anti-Rh(D) antibody clonechains were as follows.

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH10 is

GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGGTTCACCTTCAGTAGGAATGGCATGCACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGGAGTGGGTGGCATTTATATGGTTTGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGACGACACGGCTGTGTATTACTGTGCGAGAGAGGAGGCTCTGTTTCGGGGACTTACTCGGTGGTCCTACGGCATGGACGTCTGGGGCCAAGGGACCACGGTCAGCGTCTCCTCA (SEQ ID NO: 182).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH16 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGGTTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAGGGGGCTGGAGTGGGTGGCTCTTATATGGTACGATGGAGGTAACAAAGAGTATGCAGACTCCGTGAAGGGCCGCTTCAGCATCTCCAGAGACAACTCCAAGAACACTCTGTATCTGCAAGTGAACAGCCTGAGAGCCGACGACACGGCTGTCTATTACTGTGCGAGAGACCAGAGAGCAGCAGCGGGTATCTTTTATTATTCCCGTATGGACGTCTGGGGCCAAGGGA CCACGGTCACCGTCTCCTCA(SEQ ID NO: 183).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH17 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCTTGGTCCAGCCGGGGGGGTCCCTGAGACTCTCCTGTGGTGCCTCTGGAATCCCCTTTGTTTCCTCTTGGATGGCCTGGGTCCGCCAGGCCCCAGGGAAGGGGCTGGAGTGGGTGGCCAACATAAAACAAGATGGAAGTAAGAAAAACTATGTGGACTCTGTGGAGGGCCGATTCACCATCTCCAGAGACAACGCGAAGAACTCACTTTATCTGCAAATGGACAGCCTGAGAGCCGAGGACACGCGGATATATTACTGTGCGCGAGATTCACTTACTTGTTTTGACTACTGGGGCCAGGGAGCCCTGGTCACCGTCTCCTCA (SEQ ID NO: 184).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH18 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGGAGCTATGCTATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGCTACAGCATATGATGGAAAAAATAAATACTACGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCATGAACACGCTGTTTCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTTTTACTGTGCGAGAGGCGGATTTTACTATGATAGTAGTGGTTATTACGGCTTGAGGCACTACTTTGACTCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA (SEQ ID NO: 185).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH20 is

GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGAAGTTATGCTATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCGGTTATATCATATGATGGAAGTACTATATACTACGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGCCAATTCCAAGAACACGCTGTTTCTGCAAATGAACAGCCTCAGAACTGAGGACACGGCTGTATATTACTGTACGAGAGGGGGGTTTTACTATGACAGTAGTGGTTATTACGGGTTGAGGCACTACTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCA (SEQ ID NO: 186).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH24 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGCCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGCGTCTGGATTCAGCCTCAGGAGCTATGGCATGCACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGGAGTGGGTGGCAGATATATGGTTTGATGGAAGTAATAAAGATTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGTTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTATTGTGCGAGAGATTGGAGGGTGCGGGCCTTTAGTAGTGGCTGGTTAAGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCA (SEQ ID NO: 187).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH25 is

GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCGCCTGTGCAGCGTCTGGATTCAGCTTCAGGAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAGGGGGCTGGAGTGGGTGGCATTTACATGGTTTGATGGAAGCAATAAATATTATGTAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGGAAATGAACAGCCTGAGAGTCGATGACACGGCTGTATATTACTGTGCGAGAGAGGCGCCTATGCTTCGCGGAATTAGCAGATACTACTACGCGATGGACGTCTGGGGCCCAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 188).

The nucleotide sequence encoding the heavy chain of each of anti-Rh(D)antibody clones SH28, SH50, and SH53 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGGGGTCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGCGGCGTCTGGATTCACCTTCAATAGTTATGCCATGTACTGGGTCCGCCAGCCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGCTATATGGTATGATGGAAGTAATAAAGAATATGCAGATTTTGTGAAGGGCCGCTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTCTCTGCAAATGAACAGCCTGAGAGACGAGGACACGGCTGTGTATTACTGTGCGAGAGAGGCGAATCTCCTCCGTGGCTGGTCTCGATACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 189).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH32 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGAAGCGTCTAAATTCACCCTCTACAATTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCATTTATATGGTTTGATGGAAGTAATAAATACTATGAAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAACTATCTAAGAAGGTGGCACTTTCTAGGTATTACTACTATATGGACGTCTGGGGCCAGGGGACCACGGTCACTGTCTCGTCA (SEQ ID NO: 190).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH37 is

GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGTGTCTGGATTCACCCTAACTAATTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCACATGTCTGGTATGATGGAAGTAAAACAGAATACGCAGACTCCGTCAAGGGCCGATTCGCCGTCTCCAGAGACAAATCCAAGAACACACTGTTTCTGCAAATGAACAGCCTGACAGCCGAGGACACGGCTATTTATTACTGTGCGAGAGAGAGGAGAGAGAAAGTCTATATATTGTTCTACTCGTGGCTCGACCGCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA (SEQ ID NO: 191).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH39 is

GAGGTGCAGCTGCTCGAGCAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGACTGGAGTGGGTGGCAGTTATATGGTTTGATGGAAGTAATAAGGAATATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTACAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAGAAGTGGTTCGGGGAGTTATCTTATGGTCTCGGAAGTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA (SEQ ID NO: 192).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH44 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGCCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGCGTCTGGATTCAGCCTCAGGAGCTATGGCATGCACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGGAGTGGGTGGCAGATATATGGTTTGATGGAAGTAATAAAGATTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGTTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGATACGGCTGTGTATTATTGTGCGAGAGATTGGAGGGTGCGGGCCTTTAGTAGTGGCTGGTTAAGTGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCA (SEQ ID NO: 193).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH47 is

GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGCGACTCTCTTGTGCAGCCTCTGGATTCAGCTTCAGTAACTATGCTATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTACATCATTTGATGGAAGCATTAAAGACTACGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACACTATATCTGCAAATGAACAGCCTGAGAGATGAGGACACGGCTGTATATTACTGTGCGAGAGAGCGGGGGATGATAGTCGTGGTCCGTCGCAGAAATGCTTTTGATATTTGGGGCCAAGGGA CAATGGTCACCGTCTCTTCA(SEQ ID NO: 194).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH54 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGGTTCACCTTCAGTAGGAATGGCATGCACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGGAGTGGGTGGCATTTATATGGTTTGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGACGACACGGCTGTGTATTACTGTGCGAGAGAGGAGGCTCTGTTTCGGGGACTTACTCGGTGGTCCTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCAGCGTCTCCTCA (SEQ ID NO: 195).

The nucleotide sequence encoding the heavy chain of anti-Rh(D) antibodyclone SH56 is

GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGGCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTGTCTACTATGATGGAAGTAACAAACACTATTCAGACTCCGTGAAGGGCCGATTCACCATCTTCAGAGACAACTCCAAGAACACGCTGTATCTACAAATGGACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAAAGAAATTTTCGGAGTGGTTATTCCCGCTACTACTACGGTATGGACGTCTGGGGCCCAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 196).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH8 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGGCTGCGTCTGTCGGAGACAGAGTCACCATCACTTGCCGGGCAAATCAGACCATCAGAACCTCTTTAAATTGGTATCAACAAAGACCTGGGAAAGCCCCTAACCTCCTGATCTATGGTGCATCCAGGTTGCATAGTGGGGTCCCATCAAGGTTTAGTGGCGGTATTTCTGGGGCAGACTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGACTTACGGTTATTCTCGAACGTTCGGCCAAGGGACCAAGG TGGATATCAAACGA (SEQID NO: 197).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH12 is

GCCGAGCTCACCCAGTCTCCATTCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATAACTTGCCGGGCAAGTCACAACATTTACAGGTCTTTAAATTGGTTTCAGCATAAACCAGGGGAAGCCCCTAAGCTCCTGGTCTATGCTGCATCCAGTCTGCAGCGTGGGGTCCCAACCAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTTCAACCTGAAGACTCTGCGACTTACTTCTGTCAACAGAGTGTCACATTCCCCTACACTTTTGGCCAGGGGACCAAGC TGGAGATCAGACGA (SEQID NO: 198).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH13 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCGAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCCTACACTTTTGGCCAGGGGACCAAGC TGGAGATCAAACGA (SEQID NO: 199).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH14 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAACATTAGGAGGTCTTTAAATTGGTATCAACACAAACCAGGGAGAGCCCCTAGACTCCTGATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGGGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAACAGTCTGCAACCTGCAGATTTTGCAACTTACTACTGTCAGCAGAGTTCCAATACCCCGTGGACGTTCGGCCATGGGACCAAGG TGGAAATCAAACGA (SEQID NO: 200).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH16 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCCTCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAACAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCAACTTTCGGCGGAGGGACCAAGG TGGAGATCAAACGA (SEQID NO: 201).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH18 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTCTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGTATTAGCATCGCTTTAAATTGGTATCAGCAGAGACCAGGGAAAGCCCCTAAGCTCCTGATGTATGCTACATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAATATTACAATAAACCTACTTTCGGCCCTGGGACCAAGGTGG ATATCAAACGA (SEQ IDNO: 202).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH20 is

GCCGAGCTCACCCAGTCTCCATTCTCCCTGTCTGCATCTGTCGGAGACAGAGTCACCATAACTTGCCGGGCAAGTCAGAGCATTAGCAGGTCTTTAAATTGGTATCAACATAAACCAGGGGAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTCTGCAGCGTGGGGTCCCACCCAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGACTTTGCGACTTACTTCTGTCAACAGAGTGTCAGAATCCCGTACAGTTTTGGCCAGGGGACCAAGC TGGAGATCAAACGA (SEQID NO: 203).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH21 is

GCCGAGCTCACCCAGTCTCCATCCTTCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGGGCATTAGGAGTTATTTAGCCTGGTATCAGCAAAAACCAGGGAAAGCCCCTAAGCTCCTAATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCGCCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGTCAACAGCTTAATAATTACCCCCCTTTCACTTTCGGCCCTGGGACCAA AGTGGATATCAAACGA(SEQ ID NO: 204).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH24 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCACCTATTTAAATTGGTATCAGCAGAGACCAGGGAAAGCCCCTAACCTCCTGATCTATGCTGCATCCACTTTGCAAAGGGGGGTCCCATCAAGGTTCACTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACACTACCCTGTGGACGTTCGGCCAAGGGACCAAGA TGGAAATCAGACGA (SEQID NO: 205).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH26 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTTTCCGAAGGTACAGTTTTGGCCAGGGGACCA AGCTGGAGATCAAACGA(SEQ ID NO: 206).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH28 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAGATCAGAACATTAGGAGGTCTTTAAATTGGTTTCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTCCAGTACCCCGTGGACGTTCGGCCGAGGGACCAAGG TGGAAATCAAACGA (SEQID NO: 207).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH30 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTTGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTCGGAGGTCTTTAAATTGGTATCAGCAGAGTCCAGGGAAAACCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCTCACTTTCGGCGGAGGGACCAAGGTGG AGATCAAACGA (SEQ IDNO: 208).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH32 is

GCCGAGCTCACTCAGGAGCCCTCACTGACTGTGTCCCCAGGAGGGACAGTCACTCTCACCTGTGCTTCCAGCACTGGAGCAGTCACCAGTCGTTACTTTCCAAACTGGTTCCAGCAGAAACCTGGCCAGGCACCCAGGGCACTGATTTATGGTTCAAACAACAAACACTCCTGGACCCCTGCCCGGTTCTCAGGCTCCCTCCTTGGGGGCAAAGCTGCCCTGACACTGTCAGGTGTGCAGCCTGAGGACGAGGCGGAGTATTACTGCCTGCTCTTCTATGCTGGTGCTTGGGCGTTCGGCGGAGGG ACCAAGCTGACCGTCCTA(SEQ ID NO: 209).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH34 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCGGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCCCCGTACACTTTTGGCCAGGGGACCA AGCTGGAGATCAAACGA(SEQ ID NO: 210).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH36 is

GCCGAGCTCACTCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAATCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCGGCTTTCGGCCCTGGGACCAAAG TGGATATCAAACGA (SEQID NO: 211).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH39 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGACCATTGGGAGGTATTTAAATTGGTATCAGCAGAGGCCAGGGAAAGCCCCCAAACTCCTGGTATATGCTGTGTCCAGTTTGCAAAGTGGGGCCCCATCAAGGTTCAGTGGCAGTGGCTCTGGGACACATTTCACTCTCACCATCACCAGTCTGCAACCTGAAGATTTTGCAACTTACTTCTGCCAACAGAGTTACAGTTCTCCTTTCACTTTTGGCCAGGGGACCAAGGT TGAGATCAAACGA (SEQID NO: 212).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH41 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAACATTAGGAGGTCTTTAAATTGGTATCAACACAAACCAGGGAGAGCCCCTAGACTCCTGATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGGGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAACAGTCTGCAACCTGCAGATTTTGCAACTTACTACTGTCAGCAGAGTTCCAATACCCCGTGGACGTTCGGCCATGGGACCAAGG TGGAAATCAAACGA (SEQID NO: 213).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH44 is

GCCGAGCTCACCCAGTCTCCATCGTCCCTGTCTGCATCTGTAGGAGACAGAGTCATCATCACTTGCCGGGCAAGTCAGACCATTCCCAGGTTCTTGAATTGGTATCAACAGAAGCCTGGAAAAGCCCCTGTTCTCCTGATTCATAGTATATCCAGTTTACAAAGTGGGGTCCCATCAAGGTTCAGTGCCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTCGCAACTTACTACTGCCAACAGAGTTACAGTAATCTCTCTTTCGGCCCTGGGACCACAGTGGA TATTAGACGA (SEQ IDNO: 214).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH46 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGTACATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAATCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGACTTACAGTTCCCCTAGCACTTTCGGCCCTGGGACCAAAG TGGATATCAAACGA (SEQID NO: 215).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH47 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAACTATTTAAATTGGTATCAGCAGAAACCAGGAAAAGCCCCTAACCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTTATCCTCGCACGTTCGGCCAAGGGACCAAGG TGGAGATCAGACGA (SEQID NO: 216).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH48 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGTACATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAATCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGACTTACAGTTCCCCTAGCACTTTCGGCCCTGGGACCAAAG TGGATATCAAACGA (SEQID NO: 217).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH49 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCGTCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCGTGGACGTTCGGCCAAGGGACCAAG GTGGAAATCAAACGA (SEQID NO: 218).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH50 is

GCCGAGCTCACCCAGTCTCCATCGTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGACAAGTCAGAGCATTGGCACCTATTTAAATTGGTATCAACAAAAACCAGGGAAAGCCCCTAAACTCCTGATCTATGCTGCATCCAATGTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCGGTGGATCTGGGACAGGTTTCTCTCTCATCATCAGCAGTCTGCAACCTGAAGATTTAGCAATTTACTACTGCCAACAGAGCTACAGTGTCCCTCCGTACAGCTTTGGCCCGGGGACCA AGCTGGAGATCAAACGA(SEQ ID NO: 219).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH51 is

GCCGAGCTCACACAGTCTCCATCCTTCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGGGCATAAGGAGTTATTTAGCCTGGTATCAGCAAAAACCAGGGAAAGCCCCTAAGCTCCTAATCTATGCTGCATCCACTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGCTTAATAATTACCCCCCTTTCACTTTCGGCCCTGGGACCAA AGTGGATATCAAACGA(SEQ ID NO: 220).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH52 is

GCCGAGCTCACACAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTATTAGCAGCAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCAGCAGGGCCACTGGCATCCCAGACAGATTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGCAGTATGGTAGCTCACCGTGGACGTTCGGCCAAGGGACC AAGGTGGAAATCAAACGA(SEQ ID NO: 221).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH54 is

GCCGAGCTCACCCAGTCTCCATCCTCCATGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGGCACTTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCGTGGACGTTCGGCCAAGGGACCAAG GTGGAAATCAAACGA (SEQID NO: 222).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH55 is

GCCGAGCTCACGCAGCCGCCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGGAAGCAGCTCCAACATCGGAAGTAAATATGTATACTGGTACCAGCAACTCCCAGGAACGGCCCCCAAACTCCTCATTTATAGTAATAATCAGCGGCCCTCAGGGGTCCCTGACCGATTCTCTGCCTTCAAGTCTGGCACCTCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTGAGGATGAGGCTAATTATTACTGCCAGTCCTATGACAGCGGCCTGAGTGGCTGGGTGTTCGGCGGCGGGACCAAGCTGACCGTCCTA (SEQ ID NO: 223).

The nucleotide sequence encoding the light chain of anti-Rh(D) antibodyclone SH56 is

GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGGTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCCAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCGCTCTCACCATCAGCAGTCTGCTACCTGAAGATTTTGCAACTTACTACTGTCAACAGGGTTACAGTACCCCTCCGTACAGTTTTGGCCAGGGGACCA AGCTGGAGATCAAACGA(SEQ ID NO: 224).

The disclosures of each and every patent, patent application andpublication cited herein are hereby incorporated herein by reference intheir entirety.

While this invention has been disclosed with reference to specificembodiments, it is apparent that other embodiments and variations ofthis invention may be devised by others skilled in the art withoutdeparting from the true spirit and scope of the invention. The appendedclaims are intended to be construed to include all such embodiments andequivalent variations.

224 1 128 PRT Homo sapiens anti-Rh(D) chain B01 1 Glu Val Gln Leu LeuGlu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg LeuSer Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr 20 25 30 Ala Met His TrpVal Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Thr AlaTyr Asp Gly Lys Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg PheThr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe 65 70 75 80 Leu Gln MetAsn Ser Leu Arg Ala Glu Asp Thr Ala Val Phe Tyr Cys 85 90 95 Ala Arg GlyGly Phe Tyr Tyr Asp Ser Ser Gly Tyr Tyr Gly Leu Arg 100 105 110 His TyrPhe Asp Ser Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 2124 PRT Homo sapiens anti-Rh(D) chain C01 2 Glu Val Gln Leu Leu Glu SerGly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser CysAla Ala Ser Gly Phe Ser Phe Ser Ser Tyr 20 25 30 Gly Met His Trp Val ArgGln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Ser Tyr AspGly His His Lys Asn Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr IleSer Arg Asp Asn Ser Lys Lys Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn SerLeu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Asn Leu Arg GlyGlu Val Thr Arg Arg Ala Ser Val Pro Phe Asp 100 105 110 Ile Trp Gly ProGly Thr Met Val Thr Val Ser Ser 115 120 3 124 PRT Homo sapiensanti-Rh(D) chain C03 3 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val ValGln His Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly PheSer Phe Ser Ser Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly LysGly Leu Glu Trp Val 35 40 45 Ser Val Ile Ser Tyr Asp Gly His His Lys AsnTyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn SerLys Lys Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Pro Glu AspThr Ala Val Tyr Tyr Cys 85 90 95 Ala Asn Leu Arg Gly Glu Val Thr Arg ArgAla Ser Val Pro Phe Asp 100 105 110 Ile Trp Gly Pro Gly Thr Met Val ThrVal Ser Ser 115 120 4 124 PRT Homo sapiens anti-Rh(D) chain C04 4 GluVal Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Thr Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Val Ile Ser Tyr Asp Gly His Asn Lys Asn Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Lys Thr Leu Tyr 65 70 7580 Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 9095 Ala Asn Leu Arg Gly Glu Val Thr Arg Arg Ala Ser Ile Pro Phe Asp 100105 110 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 5 124PRT Homo sapiens anti-Rh(D) chain C04 5 Glu Val Gln Leu Leu Glu Ser GlyGly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys AlaAla Ser Gly Phe Ser Phe Ser Ser Tyr 20 25 30 Gly Met His Trp Val Arg GlnAla Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Tyr Asp GlyThr Asn Lys Tyr Phe Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile SerArg Asp Asn Ser Lys Lys Thr Leu Tyr 65 70 75 80 Leu Gln Met Thr Ser LeuArg Pro Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Asn Leu Arg Gly GluVal Thr Arg Arg Ala Ser Val Pro Leu Asp 100 105 110 Ile Trp Gly Gln GlyThr Met Val Thr Val Ser Ser 115 120 6 124 PRT Homo sapiens anti-Rh(D)chain C08 6 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro GlyArg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe SerSer Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu GluTrp Val 35 40 45 Ala Val Ile Ser Tyr Asp Gly Thr Asn Lys Tyr Phe Ala AspSer Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Lys ThrLeu Tyr 65 70 75 80 Leu Gln Met Thr Ser Leu Arg Pro Glu Asp Thr Ala ValTyr Phe Cys 85 90 95 Ala Asn Leu Arg Gly Glu Val Thr Arg Arg Ala Ser ValPro Leu Asp 100 105 110 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser115 120 7 124 PRT Homo sapiens anti-Rh(D) chain C10 7 Glu Val Gln LeuLeu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu ArgLeu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Ser Tyr 20 25 30 Gly Met HisTrp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val IleSer Tyr Asp Gly His His Lys Asn Tyr Ala Asp Ser Val 50 55 60 Lys Gly ArgPhe Thr Ile Ser Arg Asp Asn Ser Lys Lys Thr Leu Tyr 65 70 75 80 Leu GlnMet Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala AsnLeu Arg Gly Glu Val Thr Arg Arg Ala Ser Val Pro Phe Asp 100 105 110 IleTrp Gly Pro Gly Thr Leu Val Thr Val Ser Ser 115 120 8 125 PRT Homosapiens anti-Rh(D) chain D01 8 Glu Val Gln Leu Leu Glu Ser Gly Gly GlyVal Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Val SerGly Phe Thr Phe Asn Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala ProGly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Ser AsnLys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg AspAsn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg AlaGlu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asn Gln Ile Lys LeuTrp Ser Arg Tyr Leu Tyr Tyr Phe 100 105 110 Asp Tyr Trp Gly Gln Gly ThrLeu Val Thr Val Ser Ser 115 120 125 9 125 PRT Homo sapiens anti-Rh(D)chain D03 9 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro GlyArg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe SerThr Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu GluTrp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Glu Tyr Ala AspSer Val 50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn ThrLeu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala ValTyr Tyr Cys 85 90 95 Ala Arg Glu Glu Val Val Arg Gly Val Ile Leu Trp SerArg Lys Phe 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val SerSer 115 120 125 10 126 PRT Homo sapiens anti-Rh(D) chain D04 10 Glu ValGln Leu Leu Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 SerLeu Arg Leu Ser Cys Val Ala Ser Gly Phe Ser Leu Arg Ser Tyr 20 25 30 GlyMet His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 AlaAsp Ile Trp Phe Asp Gly Ser Asn Lys Asp Tyr Ala Asp Ser Val 50 55 60 LysGly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Trp Arg Val Arg Ala Phe Ser Ser Gly Trp Leu Ser Ala 100 105110 Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 12511 127 PRT Homo sapiens anti-Rh(D) chain D05 11 Glu Val Gln Leu Leu GluGlu Ser Gly Gly Gly Val Ala Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg LeuSer Cys Val Ala Ser Gly Phe Ser Leu Arg Ser 20 25 30 Tyr Gly Met His TrpVal Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Ala Asp Ile TrpPhe Asp Gly Ser Asn Lys Asp Tyr Ala Asp Ser 50 55 60 Val Lys Gly Arg PheThr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70 75 80 Tyr Leu Gln MetAsn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 85 90 95 Cys Ala Arg AspTrp Arg Val Arg Ala Phe Ser Ser Gly Trp Leu Ser 100 105 110 Ala Phe AspIle Trp Gly Gln Gly Thr Thr Val Ser Val Ser Ser 115 120 125 12 125 PRTHomo sapiens anti-Rh(D) chain D07 12 Glu Val Gln Leu Leu Glu Ser Gly GlyGly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala ValSer Gly Phe Thr Leu Thr Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln AlaPro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala His Val Trp Tyr Asp Gly SerLys Thr Glu Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ala Val Ser ArgAsp Lys Ser Lys Asn Thr Leu Phe 65 70 75 80 Leu Gln Met Asn Ser Leu ThrAla Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Glu Arg Arg Glu LysVal Tyr Ile Leu Phe Tyr Ser Trp Leu 100 105 110 Asp Arg Trp Gly Gln GlyThr Leu Val Thr Val Ser Ser 115 120 125 13 126 PRT Homo sapiensanti-Rh(D) chain D08 13 Glu Val Gln Leu Leu Glu Glu Ser Gly Gly Gly ValVal Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser GlyPhe Thr Phe Ser Ser 20 25 30 Tyr Gly Met His Trp Val Arg Gln Ala Pro GlyArg Gly Leu Glu Trp 35 40 45 Val Ala Leu Ile Trp Tyr Asp Gly Gly Asn LysGlu Tyr Ala Asp Ser 50 55 60 Val Lys Gly Arg Phe Ser Ile Ser Arg Asp AsnSer Lys Asn Thr Leu 65 70 75 80 Tyr Leu Gln Val Asn Ser Leu Arg Ala AspAsp Thr Ala Val Tyr Tyr 85 90 95 Cys Ala Arg Asp Gln Arg Ala Ala Ala GlyIle Phe Tyr Tyr Ser Arg 100 105 110 Met Asp Val Trp Gly Gln Gly Thr ThrVal Thr Val Ser Ser 115 120 125 14 126 PRT Homo sapiens anti-Rh(D) chainD09 14 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 15 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Lys Phe Thr Leu Tyr Asn Tyr20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val35 40 45 Ala Phe Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Glu Asp Ser Val50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr TyrCys 85 90 95 Ala Arg Glu Gly Ser Lys Lys Val Ala Leu Ser Arg Tyr Tyr TyrTyr 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser115 120 125 15 126 PRT Homo sapiens anti-Rh(D) chain D10 15 Glu Val GlnLeu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser LeuArg Leu Ser Cys Glu Ala Ser Lys Phe Thr Leu Tyr Asn Tyr 20 25 30 Gly MetHis Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala PheIle Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Glu Asp Ser Val 50 55 60 Lys GlyArg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 LeuGln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 AlaArg Glu Gly Ser Lys Lys Val Ala Leu Ser Arg Tyr Tyr Tyr Tyr 100 105 110Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 16126 PRT Homo sapiens anti-Rh(D) chain D11 16 Glu Val Gln Leu Leu Glu SerGly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser CysGlu Ala Ser Lys Phe Thr Leu Tyr Asn Tyr 20 25 30 Gly Met His Trp Val ArgGln Ala Pro Gly Glu Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Trp Phe AspGly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr ValSer Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn SerLeu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Val SerLys Lys Leu Ala Leu Ser Arg Tyr Tyr Tyr Tyr 100 105 110 Met Asp Val TrpGly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 17 126 PRT Homosapiens anti-Rh(D) chain D12 17 Glu Val Gln Leu Leu Glu Ser Gly Gly GlyVal Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ala Cys Ala Ala SerGly Phe Ser Phe Arg Ser Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala ProGly Arg Gly Leu Glu Trp Val 35 40 45 Ala Phe Thr Trp Phe Asp Gly Ser AsnLys Tyr Tyr Val Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg AspAsn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Glu Met Asn Ser Leu Arg ValAsp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Ser Met Leu ArgGly Ile Ser Arg Tyr Tyr Tyr Ala 100 105 110 Met Asp Val Trp Gly Pro GlyThr Thr Val Thr Val Ser Ser 115 120 125 18 127 PRT Homo sapiensanti-Rh(D) chain D13 18 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val ValGln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly PheThr Phe Ser Thr Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly LysGly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Ser Asn Arg AspTyr Ala Glu Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys SerLys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu AspSer Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asn Val Ala Arg Gly Gly GlyGly Val Arg Tyr Lys Tyr 100 105 110 Tyr Phe Asp Tyr Trp Gly Gln Gly ThrLeu Val Thr Val Ser Ser 115 120 125 19 127 PRT Homo sapiens anti-Rh(D)chain D14 19 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro GlyGly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe SerThr Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu GluTrp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Ser Lys Arg Asp Tyr Ala GluSer Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn ThrLeu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala ValTyr Tyr Cys 85 90 95 Ala Arg Glu Asn Val Ala Arg Gly Gly Gly Gly Ile ArgTyr Lys Tyr 100 105 110 Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val ThrVal Ser Ser 115 120 125 20 125 PRT Homo sapiens anti-Rh(D) chain D15 20Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 1015 Ser Leu Arg Leu Ser Cys Val Val Ser Gly Phe Thr Phe Asn Asn Tyr 20 2530 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 4045 Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 5560 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 7075 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 8590 95 Ala Arg Glu Asn Gln Ile Lys Leu Trp Ser Arg Tyr Leu Tyr Tyr Phe100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120125 21 125 PRT Homo sapiens anti-Rh(D) chain D16 21 Glu Val Gln Leu LeuGlu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg LeuSer Cys Val Val Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30 Gly Met His TrpVal Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile TrpPhe Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg PheThr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln MetAsn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg GluAsn Gln Ile Lys Leu Trp Ser Arg Tyr Leu Tyr Tyr Phe 100 105 110 Asp TyrTrp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 22 125 PRT Homosapiens anti-Rh(D) chain D17 22 Glu Val Gln Leu Leu Glu Ser Gly Gly GlyVal Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Val SerGly Phe Thr Phe Asn Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala ProGly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Ser AsnLys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg AspAsn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg AlaGlu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asn Gln Ile Lys LeuTrp Ser Arg Tyr Leu Tyr Tyr Phe 100 105 110 Asp Tyr Trp Gly Gln Gly ThrLeu Val Thr Val Ser Ser 115 120 125 23 125 PRT Homo sapiens anti-Rh(D)chain D18 23 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro GlyArg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Val Ser Gly Phe Thr Phe AsnAsn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Ser Gly Lys Gly Leu GluTrp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Ala AspSer Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn ThrLeu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala ValTyr Tyr Cys 85 90 95 Ala Arg Glu Asn Gln Ile Lys Leu Trp Ser Arg Tyr LeuTyr Tyr Phe 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val SerSer 115 120 125 24 125 PRT Homo sapiens anti-Rh(D) chain D20 24 Glu ValGln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 SerLeu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 GlyMet His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 AlaVal Ile Trp Phe Asp Gly Ser Asn Lys Glu Tyr Ala Asp Ser Val 50 55 60 LysGly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Glu Glu Val Val Arg Gly Val Ile Leu Trp Ser Arg Lys Phe 100 105110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 25126 PRT Homo sapiens anti-Rh(D) chain D30 25 Glu Val Gln Leu Leu Glu SerGly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser CysAla Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Gly Met Arg Trp Val ArgGln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Val Tyr Tyr AspGly Ser Asn Lys His Tyr Ser Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr IleSer Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asp SerLeu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Arg AsnPhe Arg Ser Gly Tyr Ser Arg Tyr Tyr Tyr Gly 100 105 110 Met Asp Val TrpGly Pro Gly Thr Thr Val Thr Val Ser Ser 115 120 125 26 126 PRT Homosapiens anti-Rh(D) chain D31 26 Glu Val Gln Leu Leu Glu Ser Gly Gly GlyVal Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala SerGly Phe Thr Phe Ser Ser Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala ProGly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Val Tyr Tyr Asp Gly Ser AsnLys His Tyr Ser Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg AspAsn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asp Ser Leu Arg AlaGlu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Arg Asn Phe Arg SerGly Tyr Ser Arg Tyr Tyr Tyr Gly 100 105 110 Met Asp Val Trp Gly Pro GlyThr Thr Val Thr Val Ser Ser 115 120 125 27 127 PRT Homo sapiensanti-Rh(D) chain E01is 27 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly LeuVal Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser GlyPhe Thr Phe Ser Ser Tyr 20 25 30 Ser Met His Trp Val Arg Gln Ala Pro GlyLys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Asn Ser Asn Thr Tyr IleTyr Tyr Ala Asp Ala Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp AsnAla Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala GluAsp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Ser Arg Tyr Ser Asn PheLeu Arg Trp Val Arg Ser Asp 100 105 110 Gly Met Asp Val Trp Gly Gln GlyThr Thr Val Ile Val Ser Ser 115 120 125 28 131 PRT Homo sapiensanti-Rh(D) chain E03 28 Glu Val Gln Leu Leu Glu Ser Gly Val Glu Ser GlyGly Gly Leu Val 1 5 10 15 Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys AlaAla Ser Gly Phe Thr 20 25 30 Phe Ser Ser Tyr Ser Met His Trp Val Arg GlnGly Pro Gly Lys Gly 35 40 45 Leu Glu Trp Val Ser Ser Ile Ser Asn Ser AsnThr Tyr Ile Tyr Tyr 50 55 60 Ala Asp Ala Val Lys Gly Arg Phe Thr Ile SerArg Asp Asn Ala Lys 65 70 75 80 Asn Ser Leu Tyr Leu Gln Met Asn Ser LeuArg Ala Glu His Thr Ala 85 90 95 Val Tyr Tyr Cys Ala Arg Asp Ser Arg TyrSer Asn Phe Leu Arg Trp 100 105 110 Val Arg Ser Asp Gly Met Asp Val TrpGly Gln Gly Thr Thr Val Ile 115 120 125 Val Ser Ser 130 29 107 PRT Homosapiens anti-Rh(D) chain F01 29 Ala Glu Leu Thr Gln Ser Pro Ser Ser LeuSer Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala SerGln Gly Phe Arg Asn Asp Leu 20 25 30 Gly Trp Tyr Gln Gln Lys Pro Gly LysAla Pro Lys Arg Leu Ile Tyr 35 40 45 Ala Thr Ser Ser Leu Gln Ser Gly ValPro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu ThrIle Asn Ser Leu Gln Pro Glu 65 70 75 80 Asp Ser Ala Thr Tyr Tyr Cys LeuGln His Asn Ser Phe Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val GluIle Lys Arg 100 105 30 112 PRT Homo sapiens anti-Rh(D) chain G01 30 AlaGlu Leu Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly Glu 1 5 10 15Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser Ser 20 25 30Gly Phe Asn Phe Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro 35 40 45Gln Leu Leu Ile Tyr Met Gly Ser Asn Arg Ala Ser Gly Val Pro Asp 50 55 60Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Asn 65 70 7580 Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala Leu 85 9095 Gln Phe Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100105 110 31 108 PRT Homo sapiens anti-Rh(D) chain H01 31 Ala Glu Leu ThrGln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val ThrIle Thr Cys Arg Ala Ser Gln Gly Ile Thr Ser Tyr Leu 20 25 30 Ala Trp TyrGln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala SerThr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser GlyThr Glu Phe Thr Leu Thr Ile Ala Ser Leu Gln Pro Asp 65 70 75 80 Asp PheAla Thr Tyr Tyr Cys Gln Gln Leu Asn Asn Tyr Pro Pro Phe 85 90 95 Thr PheGly Pro Gly Thr Lys Val Asp Ile Lys Arg 100 105 32 108 PRT Homo sapiensanti-Rh(D) chain I01 32 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln SerIle Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile SerSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln SerTyr Ser Thr Pro Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu IleLys Arg 100 105 33 107 PRT Homo sapiens anti-Rh(D) chain I02 33 Ala GluLeu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 ArgVal Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30 AsnTrp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 AlaAla Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 GlySer Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Leu Trp Thr 85 90 95Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 34 107 PRT Homosapiens anti-Rh(D) chain I03 34 Ala Glu Leu Thr Gln Ser Pro Ser Ser LeuSer Ala Ser Val Ala Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Thr SerArg Asn Ile Asn Arg Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly LysAla Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly ValPro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu ThrIle Thr Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys GlnGln Ser Tyr Ser Thr Pro Phe Thr 85 90 95 Phe Gly Pro Gly Thr Lys Val AspLeu Lys Arg 100 105 35 107 PRT Homo sapiens anti-Rh(D) chain I04 35 AlaGlu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Arg Arg Ser Leu 20 25 30Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn Thr Pro Trp Thr 85 9095 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 36 107 PRT Homosapiens anti-Rh(D) chain I05 36 Ala Glu Leu Thr Gln Ser Pro Ser Ser LeuSer Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala SerGln Ser Ile Arg Arg Tyr Leu 20 25 30 Asn Trp Tyr Gln His Lys Pro Gly LysAla Pro Lys Leu Leu Ile Phe 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly ValPro Ser Arg Phe Thr Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu ThrIle Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys GlnGln Ser Tyr Ser Thr Pro Gln Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val GluIle Lys Arg 100 105 37 107 PRT Homo sapiens anti-Rh(D) chain I06 37 AlaGlu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Ile Thr 85 9095 Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg 100 105 38 107 PRT Homosapiens anti-Rh(D) chain I07 38 Ala Glu Leu Thr Gln Ser Pro Ser Ser LeuSer Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala SerGln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly LysAla Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly ValPro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu ThrIle Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys GlnGln Ser Tyr Ser Thr Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val GluIle Lys Arg 100 105 39 107 PRT Homo sapiens anti-Rh(D) chain I08 39 AlaGlu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Arg Thr 85 9095 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100 105 40 107 PRT Homosapiens anti-Rh(D) chain I09 40 Ala Glu Leu Thr Gln Ser Pro Ser Ser LeuSer Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala SerGln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly LysAla Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly ValPro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Ser Thr Leu ThrIle Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys GlnGln Leu Asn Ser Tyr Pro Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Lys Leu GluIle Lys Arg 100 105 41 108 PRT Homo sapiens anti-Rh(D) chain I10 41 AlaGlu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Ser Ser Tyr Leu 20 25 30Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Ala Ala Ser Ser Leu Gln Ser Gly Val Leu Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro Tyr 85 9095 Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 42 103 PRTHomo sapiens anti-Rh(D) chain I11 42 Ala Glu Leu Thr Gln Ser Pro Ser SerLeu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg AlaSer Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro GlyLys Ala Pro Thr Leu Leu Ile Asn 35 40 45 Ala Ala Ser Ser Leu Gln Ser GlyVal Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr LeuThr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Ile Tyr Tyr CysGln Gln Arg Glu Thr Phe Gly Gln Gly 85 90 95 Thr Lys Leu Glu Ile Lys Arg100 43 108 PRT Homo sapiens anti-Rh(D) chain I12 43 Ala Glu Leu Thr GlnSer Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr IleThr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr GlnGln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser SerLeu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly ThrAsp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe AlaThr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro Tyr 85 90 95 Thr Phe GlyGln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 44 107 PRT Homo sapiensanti-Rh(D) chain I13 44 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln SerIle Ser Arg Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile SerSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln SerTyr Gly Thr Pro His Ser 85 90 95 Phe Gly Arg Gly Thr Lys Leu Glu Ile LysArg 100 105 45 107 PRT Homo sapiens anti-Rh(D) chain I15 45 Ala Glu LeuThr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg ValThr Ile Thr Cys Arg Ala Asn Gln Asn Ile Arg Arg Ser Leu 20 25 30 Asn TrpTyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 40 45 Ala AlaSer Thr Leu Gln Gly Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly SerGly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Leu Ala 65 70 75 80 AspPhe Ala Thr Tyr Tyr Cys Gln Gln Thr Ser Ala Thr Pro Trp Thr 85 90 95 PheGly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 46 107 PRT Homo sapiensanti-Rh(D) chain I16 46 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Pro AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln ThrIle Gly Phe Asn Leu 20 25 30 Asn Trp Tyr Gln Gln Thr Ser Gly Lys Pro ProLys Leu Leu Ile Tyr 35 40 45 Gly Val Ser Lys Leu Gln Asn Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile SerSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln ThrAsn Asp Ala Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Val ArgArg 100 105 47 106 PRT Homo sapiens anti-Rh(D) chain J01 47 Ala Glu LeuGln Asp Pro Val Val Ser Val Ala Leu Gly Gln Thr Val 1 5 10 15 Arg IleThr Cys Gln Gly Asp Gly Leu Arg Ser Tyr Tyr Ala Ser Trp 20 25 30 Tyr GlnGln Lys Pro Gly Gln Ala Pro Lys Leu Val Met Tyr Gly Arg 35 40 45 Asn AsnArg Pro Ser Gly Ile Pro Gly Arg Phe Ser Gly Ser Ser Ser 50 55 60 Gly GlnThr Ala Ala Leu Thr Ile Thr Gly Thr Gln Ala Glu Asp Glu 65 70 75 80 AlaAsp Tyr Tyr Cys Gln Ser Arg Ala Thr Ser Gly Asn Pro Val Val 85 90 95 PheGly Gly Gly Thr Lys Leu Thr Val Leu 100 105 48 106 PRT Homo sapiensanti-Rh(D) chain J02 48 Ala Glu Leu Gln Asp Pro Val Val Ser Val Ala LeuGly Gln Thr Val 1 5 10 15 Arg Ile Thr Cys Gln Gly Asp Gly Leu Arg SerTyr Tyr Ala Ser Trp 20 25 30 Tyr Gln Gln Lys Pro Gly Gln Ala Pro Lys LeuVal Met Tyr Gly Arg 35 40 45 Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg PheSer Gly Ser Ser Ser 50 55 60 Gly Gln Thr Ala Ala Leu Thr Ile Thr Gly ThrGln Ala Glu Asp Glu 65 70 75 80 Ala Asp Tyr Tyr Cys Gln Ser Arg Ala ThrSer Gly Asn Pro Val Val 85 90 95 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu100 105 49 104 PRT Homo sapiens anti-Rh(D) chain J04 49 Ala Glu Leu GlnAsp Pro Val Val Ser Val Ala Leu Gly Gln Thr Val 1 5 10 15 Arg Ile ThrCys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp 20 25 30 Tyr Gln GlnLys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys 35 40 45 Asn Ser ArgPro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser 50 55 60 Gly Asn ThrAla Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu 65 70 75 80 Ala AspTyr Tyr Cys Ser Ser Arg Gly Ser Pro His Val Ala Phe Gly 85 90 95 Gly GlyThr Lys Leu Thr Val Leu 100 50 106 PRT Homo sapiens anti-Rh(D) chain J0550 Ala Glu Leu Gln Asp Pro Val Val Ser Val Ala Leu Gly Gln Thr Val 1 510 15 Lys Ile Thr Cys Gln Gly Asp Ser Leu Arg Lys Tyr Tyr Ala Ser Trp 2025 30 Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Phe Tyr Ala Arg 3540 45 Asn Ser Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Asn Ser 5055 60 Gly Thr Thr Ala Ser Leu Thr Ile Ala Gly Ala Arg Ala Glu Asp Glu 6570 75 80 Ala Asp Tyr Tyr Cys His Ser Arg Asp Ser Asn Gly His His Arg Val85 90 95 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 51 108 PRT Homosapiens anti-Rh(D) chain K01 51 Ala Glu Leu Thr Gln Glu Pro Ser Leu ThrVal Ser Pro Gly Gly Thr 1 5 10 15 Val Thr Leu Thr Cys Ala Ser Ser ThrGly Ala Val Thr Ser Arg Tyr 20 25 30 Phe Pro Asn Trp Phe Gln Gln Lys ProGly Gln Ala Pro Arg Pro Leu 35 40 45 Ile Tyr Ser Ala Ser Asn Lys His SerTrp Thr Pro Ala Arg Phe Ser 50 55 60 Gly Ser Leu Leu Gly Gly Lys Ala AlaLeu Thr Leu Ser Gly Val Gln 65 70 75 80 Pro Glu Asp Glu Ala Glu Tyr TyrCys Leu Leu Tyr Tyr Ser Gly Ala 85 90 95 Trp Val Phe Gly Gly Gly Thr LysLeu Thr Val Leu 100 105 52 108 PRT Homo sapiens anti-Rh(D) chain K02 52Ala Glu Leu Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr 1 5 1015 Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Arg Tyr 20 2530 Phe Pro Asn Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Leu 35 4045 Ile Tyr Ser Ala Ser Asn Lys His Ser Trp Thr Pro Ala Arg Phe Ser 50 5560 Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln 65 7075 80 Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Leu Leu Tyr Tyr Ser Gly Ala 8590 95 Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 53 108 PRTHomo sapiens anti-Rh(D) chain K03 53 Ala Glu Leu Thr Gln Pro Pro Ser LeuThr Val Ser Pro Gly Gly Thr 1 5 10 15 Val Thr Leu Thr Cys Ala Ser SerThr Gly Ala Val Thr Ser Arg Tyr 20 25 30 Phe Pro Asn Trp Phe Gln Gln LysPro Gly Gln Ala Pro Arg Ala Leu 35 40 45 Ile Tyr Gly Ser Asn Asn Lys HisSer Trp Thr Pro Ala Arg Phe Ser 50 55 60 Gly Ser Leu Leu Gly Gly Lys AlaAla Leu Thr Leu Ser Gly Val Gln 65 70 75 80 Pro Glu Asp Glu Ala Glu TyrTyr Cys Leu Leu Phe Tyr Ala Gly Ala 85 90 95 Trp Ala Phe Gly Gly Trp ThrLys Leu Thr Val Leu 100 105 54 109 PRT Homo sapiens anti-Rh(D) chain L0154 Ala Glu Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg 1 510 15 Val Thr Ile Ser Cys Ser Gly Gly Ser Ser Asn Ile Ala Ser Asn Thr 2025 30 Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 3540 45 Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 5055 60 Ser Lys Ser Gly Thr Ser Ala Thr Leu Val Ile Thr Gly Leu Gln Thr 6570 75 80 Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp His Ser Arg Ser85 90 95 Gly Ala Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 55109 PRT Homo sapiens anti-Rh(D) chain L03 55 Ala Glu Leu Thr Gln Pro ProSer Ala Ser Gly Thr Pro Gly Gln Arg 1 5 10 15 Val Thr Ile Ser Cys SerGly Ser Ser Ser Asn Ile Gly Asn Asn His 20 25 30 Val Ser Trp Tyr Gln GlnLeu Pro Gly Met Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Asn Gly Gln ArgPro Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Lys Ser Gly Thr SerAla Ser Leu Ala Ile Ser Gly Leu Gln Ser 65 70 75 80 Glu Asp Glu Ala AspTyr Tyr Cys Ala Ala Trp His Asp Ser Leu Tyr 85 90 95 Gly Pro Val Phe GlyGly Gly Thr Lys Leu Thr Val Leu 100 105 56 109 PRT Homo sapiensanti-Rh(D) chain L04 56 Ala Glu Leu Thr Gln Pro Pro Ser Ala Ser Gly ThrPro Gly Gln Arg 1 5 10 15 Val Ser Ile Ser Cys Ser Gly Ser Ser Ser AsnIle Gly Ser Asn Thr 20 25 30 Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr AlaPro Lys Leu Leu Ile 35 40 45 Ser Thr Asn Asn Gln Gly Pro Ser Gly Val ProAsp Arg Phe Ser Gly 50 55 60 Ser Lys Ser Gly Thr Ser Ser Ser Leu Ala IleSer Gly Leu Arg Ser 65 70 75 80 Glu Ala Glu Asp Asp Tyr Tyr Cys Ala AlaTrp Asp Asp Thr Leu Asn 85 90 95 Gly Val Val Phe Gly Gly Gly Thr Lys LeuThr Val Leu 100 105 57 109 PRT Homo sapiens anti-Rh(D) chain L05 57 AlaGlu Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Leu Arg 1 5 10 15Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Ile 20 25 30Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 35 40 45Phe Ser Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55 60Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser 65 70 7580 Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Trp Asp Asp Ser Leu Asn 85 9095 Gly Arg Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 58 109PRT Homo sapiens anti-Rh(D) chain M01 58 Ala Glu Leu Thr Gln Pro Pro SerAla Ser Gly Thr Pro Gly Gln Arg 1 5 10 15 Val Thr Ile Ser Cys Ser GlySer Asn Phe Asn Ile Gly Ser Asn Tyr 20 25 30 Val Phe Trp Tyr Gln His ValPro Gly Thr Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asn Asn Asn Gln Arg ProSer Gly Val Pro Asp Arg Leu Ser Gly 50 55 60 Ser Lys Ser Gly Ala Ser AlaSer Leu Ala Ile Asn Gly Leu Arg Ser 65 70 75 80 Asp Asp Glu Ala Asp TyrTyr Cys Thr Gly Trp Asp Asp Arg Leu Ser 85 90 95 Gly Leu Ile Phe Gly GlyGly Pro Lys Val Thr Val Leu 100 105 59 109 PRT Homo sapiens anti-Rh(D)chain M02 59 Ala Glu Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly GlnArg 1 5 10 15 Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly SerAsn Tyr 20 25 30 Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys LeuLeu Ile 35 40 45 Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg PheSer Gly 50 55 60 Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly LeuArg Ser 65 70 75 80 Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp AspSer Leu Ser 85 90 95 Gly Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu100 105 60 110 PRT Homo sapiens anti-Rh(D) chain M03 60 Ala Glu Leu ThrGln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg 1 5 10 15 Val Thr IleSer Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr 20 25 30 Val Tyr TrpTyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Arg AsnAsn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Lys SerGly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg Ser 65 70 75 80 Glu AlaGlu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Ser 85 90 95 Ala ValVal Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Leu 100 105 110 61 109 PRTHomo sapiens anti-Rh(D) chain N01 61 Ala Glu Leu Thr Gln Pro Pro Ser ValSer Ala Ala Pro Gly Gln Lys 1 5 10 15 Val Thr Ile Ser Cys Ser Gly SerSer Ser Asn Ile Asp Ser Asn Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Leu ProGly Thr Ala Pro Lys Leu Leu Ile 35 40 45 Phe Asp Asn Tyr Arg Arg Pro SerGly Ile Pro Asp Arg Phe Ser Gly 50 55 60 Ser Lys Ser Gly Thr Ser Ala ThrLeu Gly Ile Thr Gly Leu Gln Thr 65 70 75 80 Gly Asp Glu Ala Asp Tyr TyrCys Ala Thr Trp Asp Asp Ser Leu Asn 85 90 95 Gly Arg Val Phe Gly Gly GlyThr Lys Leu Thr Val Leu 100 105 62 114 PRT Homo sapiens anti-Rh(D) chainN02 62 Ala Glu Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys 15 10 15 Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr20 25 30 Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile35 40 45 Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly50 55 60 Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr65 70 75 80 Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser LeuSer 85 90 95 Ala Gly Arg Val Arg Arg Met Phe Gly Gly Gly Thr Lys Leu ThrVal 100 105 110 Leu Gly 63 110 PRT Homo sapiens anti-Rh(D) chain O01 63Ala Glu Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln Arg 1 5 1015 Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Pro Tyr 20 2530 Gly Val His Trp Tyr Gln Gln Phe Pro Gly Thr Ala Pro Lys Leu Val 35 4045 Ile Tyr Asn Asp Asn Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 5560 Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 7075 80 Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser Leu 8590 95 Ser Gly Arg Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105110 64 112 PRT Homo sapiens anti-Rh(D) chain O02 64 Ala Glu Leu Thr GlnPro Pro Ser Val Ser Gly Ala Pro Gly Gln Thr 1 5 10 15 Val Thr Ile SerCys Thr Gly Ser Ser Ser Ser Ile Gly Ala Arg Tyr 20 25 30 Asp Val His TrpTyr Gln His Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Gly AsnHis Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys SerGly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ala Glu AspGlu Ala Glu Tyr Tyr Cys Gln Ser Tyr Asp Asn Ser Leu 85 90 95 Ser Gly SerSer Val Phe Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110 65 110PRT Homo sapiens anti-Rh(D) chain O03 65 Ala Glu Leu Thr Gln Pro Pro SerGly Ala Pro Gly Gln Thr Val Thr 1 5 10 15 Ile Ser Cys Thr Gly Ser SerSer Asn Ile Gly Ala Gly Tyr Asp Val 20 25 30 His Trp Tyr Gln Gln Leu ProGly Thr Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Gly Asn Ser Asn Arg Pro SerGly Val Pro Asp Arg Phe Ser Gly Ser 50 55 60 Lys Ser Gly Thr Ser Ala SerLeu Ala Ile Thr Gly Leu Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr TyrCys Gln Ser Tyr Asp Ser Ser Leu Ser Gly 85 90 95 Pro Tyr Val Val Phe GlyGly Gly Thr Lys Leu Thr Val Leu 100 105 110 66 108 PRT Homo sapiensanti-Rh(D) chain P01 66 Ala Glu Leu Thr Gln Pro Pro Ser Val Ser Val AlaPro Arg Gln Thr 1 5 10 15 Ala Arg Ile Thr Cys Gly Gly Asp Lys Ile GlySer Asn Thr Val His 20 25 30 Trp Tyr Arg Gln Met Ser Gly Gln Ala Pro ValLeu Val Ile Tyr Glu 35 40 45 Asp Lys Lys Arg Pro Pro Gly Ile Pro Glu ArgPhe Ser Gly Ser Thr 50 55 60 Ser Gly Thr Thr Ala Thr Leu Ser Ile Ser GlyAla Gln Val Glu Asp 65 70 75 80 Glu Ala Asp Tyr Tyr Cys Tyr Ser Arg AspAsn Ser Gly Asp Gln Arg 85 90 95 Arg Val Phe Gly Ala Gly Thr Lys Leu ThrVal Leu 100 105 67 110 PRT Homo sapiens anti-Rh(D) chain Q01 67 Ala GluLeu Thr Gln Pro Pro Ser Ala Thr Ala Ser Leu Gly Gly Ser 1 5 10 15 ValLys Leu Thr Cys Ile Leu Gln Ser Gly His Arg Asn Tyr Ala Val 20 25 30 AlaTrp His His Gln Glu Ala Gly Lys Gly Pro Arg Phe Leu Met Thr 35 40 45 ValThr Asn Asp Gly Arg His Ile Lys Gly Asp Gly Ile Pro Asp Arg 50 55 60 PheSer Gly Ser Ala Ser Gly Ala Glu Arg Tyr Leu Ser Ile Ser Gly 65 70 75 80Leu Gln Ser Glu Asp Glu Gly Asp Tyr Tyr Cys Gln Thr Trp Gly Thr 85 90 95Gly Met His Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110 68108 PRT Homo sapiens anti-Rh(D) chain R01 68 Ala Glu Leu Thr Gln Pro ProSer Ala Ser Gly Ser Pro Gly Gln Ser 1 5 10 15 Val Thr Ile Ser Cys ThrGly Ala Ser Ser Asp Val Gly Ala Tyr Lys 20 25 30 His Val Ser Trp Tyr GlnGln His Pro Gly Lys Ala Pro Lys Leu Leu 35 40 45 Thr His Glu Gly Thr LysArg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly AsnThr Ala Ser Leu Thr Val Ser Gly Leu Gln 65 70 75 80 Ala Glu Asp Glu AlaAsp Tyr Tyr Cys Ser Ser Phe Ala Gly Asn Ser 85 90 95 Val Ile Phe Gly GlyGly Thr Lys Leu Thr Val Leu 100 105 69 104 PRT Homo sapiens anti-Rh(D)chain S01 69 Ala Glu Leu Thr Gln Pro Pro Ser Val Ser Gly Ser Pro Gly GlnSer 1 5 10 15 Ile Thr Ile Ser Cys Ser Asp Val Gly Asn Tyr Asn Leu ValSer Trp 20 25 30 Tyr Gln Gln Tyr Pro Gly Lys Ala Pro Lys Leu Ile Ile TyrGlu Gly 35 40 45 Ser Lys Arg Pro Ser Gly Val Ser Ser Arg Phe Ser Gly SerArg Ser 50 55 60 Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala GluAsp Glu 65 70 75 80 Ala Asp Tyr His Cys Cys Ser Tyr Ala Ile Ser Ser ArgIle Phe Gly 85 90 95 Gly Gly Thr Lys Leu Thr Val Leu 100 70 384 DNA Homosapiens anti-Rh(D) chain B01 70 gaggtgcagc tgctcgagtc tgggggaggcgtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggatt caccttcaggagctatgcta tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagctacagcatatg atggaaaaaa taaatactac 180 gcagactccg tgaagggccg attcaccatctccagagaca attccaagaa cacgctgttt 240 ctgcaaatga acagcctgag agctgaggacacggctgtgt tttactgtgc gagaggcgga 300 ttttactatg atagtagtgg ttattacggcttgaggcact actttgactc ctggggccag 360 ggaaccctgg tcaccgtctc ctca 384 71372 DNA Homo sapiens anti-Rh(D) chain C03 71 gaggtgcagc tgctcgagtctgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggattctccttcagt agctatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtgggtgtcagtt atatcatatg atggacatca taaaaactat 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccaagaa aacgctgtac 240 ctgcaaatga acagcctgagacctgaggac acggctgtat attactgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgcgtctgttccc tttgatatct ggggcccagg gacaatggtc 360 accgtctctt ca 372 72 372DNA Homo sapiens anti-Rh(D) chain C01 72 gaggtgcagc tgctcgagtcggggggaggt gtggtccagc atgggaggtc cctgagactg 60 tcctgtgcag cctctggattctccttcagt agctatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtgggtgtcagtt atatcatatg atggacatca taaaaactat 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccaagaa aacgctgtac 240 ctgcaaatga acagcctgagacctgaggac acggctgtat attactgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgcgtctgttccc tttgatatat ggggcccagg gacaatggtc 360 accgtgtctt ca 372 73 372DNA Homo sapiens anti-Rh(D) chain C04 73 gaggtgcagc tgctcgagtctgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggattctccttcagt acctatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtgggtgtcagtt atatcatatg atggacataa taaaaactat 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccaagaa aacgctgtac 240 ctgcaaatga acagcctgagacctgaggac acggctgtgt attactgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgcgtctattcct tttgatatct ggggccaagg gacaatggtc 360 accgtctctt ca 372 74 372DNA Homo sapiens anti-Rh(D) chain C05 74 gaggtgcagc tgctcgagtcggggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggattcagcttcagt agttatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtgggtggcagtt atatcgtatg atggaactaa taaatacttt 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccaagaa aacgctgtat 240 ctgcaaatga ccagcctgagacctgaggac acggctgtgt atttctgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgcgtccgtacct cttgatatct ggggccaagg gacaatggtc 360 accgtctctt ca 372 75 372DNA Homo sapiens anti-Rh(D) chain C08 75 gaggtgcagc tgctcgagtcggggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggattcagcttcagt agttatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtgggtggcagtt atatcgtatg atggaactaa taaatacttt 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccaagaa aacgctgtat 240 ctgcaaatga ccagcctgagacctgaggac acggctgtgt atttctgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgcgtctgtacct cttgatatct ggggccaagg gacaatggtc 360 accgtctctt ca 372 76 372DNA Homo sapiens anti-Rh(D) chain C10 76 gaggtgcagc tgctcgagtctgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggattctccttcagt agctatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtgggtgtcagtt atatcatatg atggacatca taaaaactat 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccaagaa aacgctgtac 240 ctgcaaatga acagcctgagacctgaggac acggctgtat attactgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgcgtctgttccc tttgatatct ggggcccagg gacattggtc 360 accgtctctt ca 372 77 375DNA Homo sapiens anti-Rh(D) chain D01 77 gaggtgcagc tgctcgagtctgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgtag tgtctggtttcaccttcaat aactatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtgggtggcagtt atttggtttg atggaagtaa taaatactat 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccaagaa cacactgtac 240 ctgcaaatga acagcctgagagccgaggac acggctgtat attactgtgc gagagagaac 300 cagataaagc tatggtcccgatacctttac tactttgatt actggggcca gggaaccctg 360 gtcaccgtct cctca 375 78375 DNA Homo sapiens anti-Rh(D) chain D03 78 gaggtgcagc tgctcgagtctgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cgtctggattcaccttcagt acctatggca tgcactgggt ccgccaggct 120 ccaggcaagg gactggagtgggtggcagtt atatggtttg atggaagtaa taaggaatat 180 gcagactccg tgaagggccgattcaccgtc tccagagaca attccaagaa cacgctgtat 240 ctacaaatga acagcctgagagccgaggac acggctgtgt attactgtgc gagagaagaa 300 gtggttcggg gagttatcttatggtctcgg aagtttgact actggggcca gggaaccctg 360 gtcaccgtct cctca 375 79378 DNA Homo sapiens anti-Rh(D) chain D04 79 gaggtgcagc tgctcgagtcggggggaggc gtggcccagc ctgggaggtc cctgagactc 60 tcctgtgtag cgtctggattcagcctcagg agctatggca tgcactgggt ccgccaggct 120 cctggcaagg ggctggagtgggtggcagat atatggtttg atggaagtaa taaagattat 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccaagaa cacgttgtat 240 cttcaaatga acagcctgagagccgaggat acggctgtgt attattgtgc gagagattgg 300 agggtgcggg cctttagtagtggctggtta agtgcttttg atatctgggg ccaagggaca 360 atggtcaccg tctcctca 37880 381 DNA Homo sapiens anti-Rh(D) chain D05 80 gaggtgcagc tgctcgaggagtctggggga ggcgtggccc agcctgggag gtccctgaga 60 ctctcctgtg tagcgtctggattcagcctc aggagctatg gcatgcactg ggtccgccag 120 gctcctggca aggggctggagtgggtggca gatatatggt ttgatggaag taataaagat 180 tatgcagact ccgtgaagggccgattcacc atctccagag acaattccaa gaacacgttg 240 tatcttcaaa tgaacagcctgagagccgag gacacggctg tgtattattg tgcgagagat 300 tggagggtgc gggcctttagtagtggctgg ttaagtgctt ttgatatctg gggccaaggg 360 accacggtca gcgtctcctc a381 81 375 DNA Homo sapiens anti-Rh(D) chain D07 81 gaggtgcagctgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcagtgtctggatt caccctaact aattatggca tgcactgggt ccgccaggct 120 ccaggcaaggggctggagtg ggtggcacat gtctggtatg atggaagtaa aacagaatat 180 gcagactccgtcaagggccg attcgccgtc tccagagaca aatccaagaa cacactgttt 240 ctgcaaatgaacagcctgac agccgaggac acggctattt attactgtgc gagagagagg 300 agagagaaagtctatatatt gttctactcg tggctcgacc gctggggcca gggaaccctg 360 gtcaccgtctcctca 375 82 378 DNA Homo sapiens anti-Rh(D) chain D08 82 gaggtgcagctgctcgagga gtctggggga ggcgtggtcc agcctgggag gtccctgaga 60 ctctcctgtgcagcgtctgg gttcaccttc agtagctatg gcatgcactg ggtccgccag 120 gctccaggcagggggctgga gtgggtggct cttatatggt acgatggagg taacaaagag 180 tatgcagactccgtgaaggg ccgcttcagc atctccagag acaattccaa gaacactctg 240 tatctgcaagtgaacagcct gagagccgac gacacggctg tctattactg tgcgagagac 300 cagagagcagcagcgggtat cttttattat tcccgtatgg acgtctgggg ccaagggacc 360 acggtcaccgtctcctca 378 83 378 DNA Homo sapiens anti-Rh(D) chain D09 83 gaggtgcagctgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgaagcgtctaaatt caccctctac aattatggca tgcactgggt ccgccaggct 120 ccaggcaaggggctggagtg ggtggcattt atatggtttg atggaagtaa taaatactat 180 gaagactccgtgaagggccg attcaccgtc tccagagaca attccaagaa cacgctgtat 240 ctgcaaatgaacagcctgag agccgaggac acggctgtgt attactgtgc gagagaagga 300 tctaagaaggtggcactttc taggtattac tattatatgg acgtctgggg ccaggggacc 360 acggtcactgtctcgtca 378 84 378 DNA Homo sapiens anti-Rh(D) chain D10 84 gaggtgcagctgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgaagcgtctaaatt caccctctac aattatggca tgcactgggt ccgccaggct 120 ccaggcaaggggctggagtg ggtggcattt atatggtttg atggaagtaa taaatactat 180 gaagactccgtgaagggccg attcaccgtc tccagagaca attccaagaa cacgctgtat 240 ctgcaaatgaacagcctgag agccgaggac acggctgtgt attactgtgc gagagaagta 300 tctaagaaggtggcactttc taggtattac tactatatgg acgtctgggg ccaggggacc 360 acggtcactgtctcctca 378 85 378 DNA Homo sapiens anti-Rh(D) chain D11 85 gaggtgcagctgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgaagcgtctaaatt caccctctac aattatggca tgcactgggt ccgccaggct 120 ccaggcgaagggctggagtg ggtggcattt atatggtttg atggaagtaa taaatactat 180 gcagactccgtgaagggccg attcaccgtc tccagagaca attccaagaa cacgctgtat 240 ctgcaaatgaacagcctgag agccgaggac acggctgtgt attactgtgc gagagaagta 300 tctaagaagctggcactttc taggtactac tactatatgg acgtctgggg ccaggggacc 360 acggtcactgtctcctca 378 86 378 DNA Homo sapiens anti-Rh(D) chain D12 86 gaggtgcagctgctcgagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60 gcctgtgcagcgtctggatt cagcttcagg agctatggca tgcactgggt ccgccaggct 120 ccaggcagggggctggagtg ggtggcattt acatggtttg atggaagcaa taaatattat 180 gtagactccgtgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240 ctggaaatgaacagcctgag agtcgatgac acggctgtat attactgtgc gagagaggcg 300 tctatgcttcgcggaattag cagatactac tacgcgatgg acgtctgggg cccagggacc 360 acggtcaccgtctcctca 378 87 381 DNA Homo sapiens anti-Rh(D) chain D13 87 gaggtgcagctgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcagcgtctggatt caccttcagt acttatggca tgcactgggt ccgccaggct 120 ccaggcaaggggctggagtg ggtggcagtt atatggtttg atggaagtaa cagagactat 180 gcagagtccgtgaagggccg attcaccatc tccagagaca agtccaagaa cacactgtat 240 ctgcaaatgaacagcctgag agccgaggac tcggctgtgt attattgtgc gagagaaaat 300 gtggctcgtggggggggggg cgttcgatac aagtactact ttgactactg gggccaggga 360 accctggtcaccgtctcctc a 381 88 381 DNA Homo sapiens anti-Rh(D) chain D14 88gaggtgcagc tgctcgagtc ggggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt acttatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatggtttg atggaagtaa gagagactat 180gcagagtccg tgaagggccg attcaccatc tccagagaca actccaagaa cacactgtat 240ctgcaaatga acagcctgag agccgaggac tcggctgtgt attactgtgc gagagaaaat 300gtggctcgtg gggggggggg cattcgatac aagtactact ttgactactg gggccaggga 360accctggtca ccgtctcctc a 381 89 375 DNA Homo sapiens anti-Rh(D) chain D1589 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgtag tgtctggatt caccttcaat aactatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atttggtttg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacactgtac 240ctgcaaatga acagcctgag agccgaggac acggctgtat attactgtgc gagagagaac 300cagataaagc tatggtcccg atacctttac tactttgact actggggcca gggaaccctg 360gtcaccgtct cctca 375 90 375 DNA Homo sapiens anti-Rh(D) chain D16 90gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgtag tgtctggttt caccttcaat aactatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atttggtttg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacactgtac 240ctgcaaatga acagcctgag agccgaggac acggctgtat attactgtgc gagagagaac 300cagataaagc tatggtcccg atacctttac tactttgact actggggcca gggaaccctg 360gtcaccgtct cctca 375 91 375 DNA Homo sapiens anti-Rh(D) chain D17 91gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgtag tgtctggttt caccttcaat aactatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atttggtttg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacactgtac 240ctgcaaatga acagcctgag agccgaggac acggctgtat attactgtgc gagagagaac 300cagataaagc tatggtcccg atacctttac tactttgact actggggcca gggaaccctg 360gtcaccgtct cctcc 375 92 375 DNA Homo sapiens anti-Rh(D) chain D18 92gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgtag tgtctggttt caccttcaat aactatggca tgcactgggt ccgccaggct 120tcaggcaagg ggttggagtg ggtggcagtt atttggtttg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacactgtac 240ctgcaaatga acagcctgag agccgaggac acggctgtat attactgtgc gagagagaac 300cagataaagc tatggtcccg atacctttac tactttgact actggggcca gggaaccctg 360gtcaccgtgt cctca 375 93 375 DNA Homo sapiens anti-Rh(D) chain D20 93gaggtgcagc tgctcgagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt acctatggca tgcactgggt ccgccaggct 120ccaggcaagg gactggagtg ggtggcagtt atatggtttg atggaagtaa taaggaatat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctacaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagagaagaa 300gtggttcggg gagttatctt atggtctcgg aagtttgact actggggcca gggaaccctg 360gtcaccgtct cctca 375 94 378 DNA Homo sapiens anti-Rh(D) chain D30 94gaggtgcagc tgctcgagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt agctatggca tgcgctgggt ccggcaggct 120ccaggcaagg ggctggagtg ggtggcagtt gtctactatg atggaagtaa caaacactat 180tcagactccg tgaagggccg attcaccatc tccagagaca actccaagaa cacgctgtat 240ctacaaatgg acagcctgag agccgaggac acggctgtgt attactgtgc gagagaaaga 300aattttcgga gtggttattc ccgctactac tacggtatgg acgtctgggg cccagggacc 360acggtcaccg tctcctca 378 95 378 DNA Homo sapiens anti-Rh(D) chain D31 95gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt agctatggca tgcactgggt ccggcaggct 120ccaggcaagg ggctggagtg ggtggcagtt gtctactatg atggaagtaa caaacactat 180tcagactccg tgaagggccg attcaccatc tccagagaca actccaagaa cacgctgtat 240ctacaaatgg acagcctgag agccgaggac acggctgtgt attactgtgc gagagaaaga 300aattttcgga gtggttattc ccgctactac tacggtatgg acgtctgggg cccagggacc 360acggtcaccg tctcctca 378 96 381 DNA Homo sapiens anti-Rh(D) chain E01 96gaggtgcagc tgctcgagtc tgggggaggc ctggtcaagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt agctatagca tgcactgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcatcc attagtaata gtaatactta catatactac 180gcagacgcag tgaagggccg attcaccatc tccagagaca acgccaagaa ctcactgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt actactgtgc gagagattct 300agatacagta atttcctccg ttgggttcgg agcgacggta tggacgtctg gggccaaggg 360accacggtca tcgtctcctc a 381 97 393 DNA Homo sapiens anti-Rh(D) chain E0397 gaggtgcagc tgctcgagtc tggggtggag tctgggggag gcctggtcaa gcctgggggg 60tccctgagac tctcctgtgc agcctctgga ttcaccttca gtagctatag catgcactgg 120gtccgccagg gtccagggaa ggggctggag tgggtctcat ccattagtaa tagtaatact 180tacatatact acgcagacgc agtgaagggc cgattcacca tctccagaga caacgccaag 240aactcactgt atctgcaaat gaacagcctg agagccgagc acacggctgt gtactactgt 300gcgagagatt ctagatacag taatttcctc cgttgggttc ggagcgacgg tatggacgtc 360tggggccaag ggaccacggt catcgtctcc tca 393 98 321 DNA Homo sapiensanti-Rh(D) chain F01 98 gccgagctca cccagtctcc atcctccctg tctgcatctgtaggagacag agtcaccatc 60 acttgccggg caagtcaggg ctttagaaat gatttaggctggtatcagca gaaaccaggg 120 aaagccccta agcgcctgat ctatgctaca tccagtttgcaaagtggggt cccatcaagg 180 ttcagcggca gtggatctgg gacagaattc actctcacaatcaacagcct gcagcctgaa 240 gattctgcaa cttattactg tctacagcat aatagtttcccgtggacgtt cggccaaggg 300 accaaggtgg aaatcaaacg a 321 99 336 DNA Homosapiens anti-Rh(D) chain G01 99 gccgagctca ctcagtctcc actctccctgcccgtcaccc ctggagagcc ggcctccatc 60 tcctgcaggt ctagtcagag cctcctgcatagtagtggat tcaacttttt ggattggtac 120 ctgcagaagc cagggcagtc tccacagctcctgatctata tgggttctaa tcgggcctcc 180 ggggtccctg acaggttcag tggcagtggatcaggcacag attttacact gaaaatcaac 240 agagtggagg ctgaggatgt tggggtttattactgcatgc aagctctaca atttcctctc 300 actttcggcg gagggaccaa ggtggagatcaaacga 336 100 324 DNA Homo sapiens anti-Rh(D) chain H01 100 gccgagctcacccagtctcc atccttcctg tctgcatctg taggagacag agtcaccatc 60 acttgccgggccagtcaggg cattacgagt tatttagcct ggtatcagca aaaaccaggg 120 aaagcccctaagctcctaat ctatgctgca tccactttgc aaagtggggt cccatcaagg 180 ttcagcggcagtggatctgg gacagaattc actctcacaa tcgccagcct gcagcctgat 240 gattttgcaacttattactg tcaacagctt aataattacc cccctttcac tttcggccct 300 gggaccaaagtggatatcaa acga 324 101 324 DNA Homo sapiens anti-Rh(D) chain I01 101gccgagctca cccagtctcc atcctcccta tctgcatctg taggagacag agtcaccatc 60acttgccggg caagtcagag cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240gattttgcaa cttactactg tcaacagagt tacagtaccc ctccgtacac ttttggccag 300gggaccaagc tggagatcaa acga 324 102 321 DNA Homo sapiens anti-Rh(D) chainI02 102 gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacagagtcaccatc 60 acttgccggg caagtcagag cattagcagc tatttaaatt ggtatcagcagaaaccaggg 120 aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggtcccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtctgcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tacagtaccc tgtggacgttcggccaaggg 300 accaaggtgg aaatcaaacg a 321 103 321 DNA Homo sapiensanti-Rh(D) chain I03 103 gccgagctca cccagtctcc atcctccctg tctgcatctgtagcggacag agtcaccatc 60 acttgccgga caagtcggaa cattaacaga tacttaaattggtatcagca gaaaccaggg 120 aaagccccta agctcctgat ttatgctgca tccagtttgcaaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcaccatcaccagtct gcaacctgaa 240 gattttgcca cttactactg tcaacagagt tacagtacccctttcacttt cggccctggg 300 accaaagtgg atctcaaacg a 321 104 321 DNA Homosapiens anti-Rh(D) chain I04 104 gccgagctca ctcagtctcc atcctccctgtctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagaa cattaggaggtctttaaatt ggtatcaaca gaaaccaggg 120 aaagccccta agctcctgat ctatgctgcatccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg gacagatttcactctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactg tcagcagagttccaataccc cgtggacgtt cggccaaggg 300 accaaggtgg aaatcaaacg a 321 105 321DNA Homo sapiens anti-Rh(D) chain I05 105 gccgagctca cccagtctccatcctccctg tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagagcattaggagg tatttaaatt ggtatcagca caaaccaggg 120 aaagccccta agctcctgatctttgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcactggca gtggatctgggacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactgtcaacagagt tacagtaccc ctcaaacgtt cggccaaggg 300 accaaggtgg aaatcaaacg a321 106 321 DNA Homo sapiens anti-Rh(D) chain I06 106 gccgagctcacccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc 60 acttgccgggcaagtcagag cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagcccctaagctcctgat ctatgccgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggcagtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaacttactactg tcaacagagt tacagtaccc cgatcacctt cggccaaggg 300 acacgactggagattaaacg a 321 107 321 DNA Homo sapiens anti-Rh(D) chain I07 107gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc 60acttgccggg caagtcagag cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240gattttgcaa cttactactg tcaacagagt tacagtaccc ctcgaacttt cggcggaggg 300accaaggtgg agatcaaacg a 321 108 321 DNA Homo sapiens anti-Rh(D) chainI08 108 gccgagctca cccagtctcc attctccctg tctgcatctg tcggagacagagtcaccata 60 acttgccggg caagtcagac cattagcagg tctttaaatt ggtatcagcataaaccaggg 120 gaagccccta agctcctgat ctatgctgca tccagtctgc agcgtggggtcccacccagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtctgcaacctgaa 240 gactttgcga cttacttctg tcaacagagt gtcagaatcc cgtacagttttggccagggg 300 accaagctgg agatcaaacg a 321 109 321 DNA Homo sapiensanti-Rh(D) chain I09 109 gccgagctca cccagtctcc atcctccctg tctgcatctgtaggagacag agtcaccatc 60 acttgccggg caagtcagag cattagcagc tatttaaattggtatcagca gaaaccaggg 120 aaagccccta agctcctgat ctatgctgca tccagtttgcaaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg gacagattcc actctcaccatcagcagtct gcaacctgaa 240 gattttgcaa cttattactg tcaacagctt aatagttacccgtacacttt tggccagggg 300 accaagctgg agatcaaacg a 321 110 324 DNA Homosapiens anti-Rh(D) chain I10 110 gccgagctca cccagtctcc atcctccctgtctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagaa cattagcagctatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta agctcctgat ctatgctgcatccagtttgc aaagtggggt cctatcaagg 180 ttcagtggca gtggatctgg gacagatttcactctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactg tcaacagagttacagtaccc ctccgtatag ttttggccag 300 gggaccaagc tggagatcaa acga 324 111309 DNA Homo sapiens anti-Rh(D) chain I11 111 gccgagctca cccagtctccatcctccctg tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagagcattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta cgctcctgatcaatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgggacagatttc actctcacca ttagcagtct gcaacctgaa 240 gatttcgcaa tttactactgtcaacagaga gaaacttttg gccaggggac caagctggag 300 atcaaacga 309 112 324DNA Homo sapiens anti-Rh(D) chain I12 112 gccgagctca cccagtctccatcctcccta tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagagcattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta agctcctgatctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgggacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactgtcaacagagt tacagtaccc ctccgtacac ttttggccag 300 gggaccaagc tggagatcaaacga 324 113 321 DNA Homo sapiens anti-Rh(D) chain I13 113 gccgagctcacccagtctcc atcctccctg tctgcctctg taggagacag agtcaccatc 60 acttgccgggcaagtcagag cattagcagg tatttaaatt ggtatcagca gaaaccaggg 120 aaagcccctaagctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggcagtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaacttactactg tcaacagagt tacggtaccc ctcacagttt tggccggggg 300 accaagctggagatcaaacg a 321 114 321 DNA Homo sapiens anti-Rh(D) chain I15 114gccgagctca cccagtctcc ttcctccctg tctgcatctg taggagacag agtcaccatc 60acttgccggg caaatcagaa cattcgtaga tctttaaatt ggtatcagca gaaaccaggg 120aaagccccta acctcctgat ctatgctgca tccacattgc aaggtggggt cccatcaagg 180ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacttgcg 240gattttgcaa cttactactg tcaacagact tccgctaccc cgtggacgtt cggccaaggg 300accaaggtgg aaatcaaacg a 321 115 321 DNA Homo sapiens anti-Rh(D) chainI16 115 gccgagctca cccagtctcc atcgtccctg cctgcatctg tgggagacagagtcaccatc 60 acttgccggg caagtcagac tattggtttt aatttaaatt ggtatcagcaaacatctggg 120 aagcccccta aactcctaat ctatggtgtt tccaagttgc aaaatggggtcccttcacgg 180 ttcagtggca gtgggtccgg gacggaattc accctcacaa tcagcagtctgcagcctgag 240 gattttgcga cttattattg tcaacagact aacgatgcgt tgtggacgttcggccaaggg 300 accaaagtgg aagtcagacg a 321 116 318 DNA Homo sapiensanti-Rh(D) chain J01 116 gccgagctcc aggaccctgt tgtgtctgtg gccttgggacagacagtcag gatcacttgc 60 caaggagacg gcctcagaag ttattatgca agctggtaccagcagaagcc gggacaggcc 120 ccgaaacttg tcatgtacgg tagaaacaac cggccctcagggatcccagg ccgattctct 180 ggctccagct cagggcagac agctgccttg accatcacggggactcaggc ggaggatgag 240 gctgactatt actgtcagtc ccgtgccacc agcggtaaccctgtggtgtt cggcggaggg 300 actaagctga ccgtcctg 318 117 318 DNA Homosapiens anti-Rh(D) chain J02 117 gccgagctcc aggaccctgt tgtgtctgtggccttgggac agacagtcag gatcacttgc 60 caaggagacg gcctcagaag ttattatgcaagctggtacc agcagaagcc gggacaggcc 120 ccgaaacttg tcatgtacgg tagaaacaaccggccctcag ggatcccaga ccgattctct 180 ggctccagct cagggcagac agctgccttgaccatcacgg ggactcaggc ggaggatgag 240 gctgactatt actgtcagtc ccgtgccaccagcggtaacc ctgtggtgtt cggcggaggg 300 actaagctga ccgtcctg 318 118 312 DNAHomo sapiens anti-Rh(D) chain J04 118 gccgagctcc aggaccctgt tgtgtctgtggccttgggac agacagtcag gatcacatgc 60 caaggagaca gcctcagaag ctattatgcaagctggtacc agcagaagcc aggacaggcc 120 cctgtacttg tcatctatgg taaaaacagccggccctcag ggatcccaga ccgattctct 180 ggctccagct caggaaacac agcttcgttgaccatcactg gggctcaggc ggaagatgag 240 gcggactatt attgtagttc gcggggcagcccccacgtgg cattcggcgg agggaccaaa 300 ctgaccgtcc tg 312 119 318 DNA Homosapiens anti-Rh(D) chain J05 119 gccgagctcc aggaccctgt tgtgtctgtggccttgggac agacagtcaa gatcacatgc 60 cagggagaca gcctcagaaa gtattatgcaagctggtacc agcagaagcc aggacaggcc 120 cctgtgcttg tcttctatgc tagaaatagccggccctcag ggatcccaga ccgattctct 180 ggctccaact caggaaccac agcttccttgaccatcgctg gggctcgggc ggaagatgag 240 gctgactatt actgtcactc ccgggacagcaatggtcacc atcgggtgtt cggcggaggg 300 accaagctga ccgtccta 318 120 324 DNAHomo sapiens anti-Rh(D) chain K01 120 gccgagctca ctcaggagcc ctcactgactgtgtccccag gagggacagt cactctcacc 60 tgtgcttcca gcactggagc agtcaccagtcgttactttc caaactggtt ccagcagaaa 120 cctggacaag cacccaggcc actgatttatagtgcaagca acaaacactc ctggacccct 180 gcccggttct caggctccct ccttgggggcaaagctgccc tgacactgtc aggtgtgcag 240 cctgaggacg aggctgagta ttactgcctgctctactata gtggtgcttg ggtgttcggc 300 ggagggacca agttgaccgt cctt 324 121324 DNA Homo sapiens anti-Rh(D) chain K02 121 gccgagctca ctcaggagccctcactgact gtgtccccag gagggacagt cactctcacc 60 tgtgcttcca gcactggagcagtcaccagt cgttactttc caaactggtt ccagcagaaa 120 cctggacaag cacccaggccactgatttat agtgcaagca acaaacactc ctggacccct 180 gcccggttct caggctccctccttgggggc aaagctgccc tgacactgtc aggtgtgcag 240 cctgaggacg aggctgagtattactgcctg ctctactata gtggtgcttg ggtgttcggc 300 ggagggacca agctgaccgtccta 324 122 324 DNA Homo sapiens anti-Rh(D) chain K03 122 gccgagctcactcagccacc ctcactgact gtgtccccag gagggacagt cactctcacc 60 tgtgcttccagcactggagc agtcaccagt cgttactttc caaactggtt ccagcagaaa 120 cctggccaggcacccagggc actgatttat ggttcaaaca acaaacactc ctggacccct 180 gcccggttctcaggctccct ccttgggggc aaagctgccc tgacactgtc aggtgtgcag 240 cctgaggacgaggcggagta ttactgcctg ctcttctatg ctggtgcttg ggcgttcggc 300 ggatggaccaagctgaccgt ccta 324 123 327 DNA Homo sapiens anti-Rh(D) chain L01 123gccgagctca cgcagccgcc ctcagcgtct gggacccccg ggcagagggt caccatctct 60tgttctggag gcagctccaa catcgcaagt aatactgtaa actggtacca gcaactccca 120ggaacggccc ccaaactcct catctatagt aataatcagc ggccctcagg ggtccctgac 180cgattctctg gctccaagtc tggcacctca gccaccctgg tcatcaccgg gctccagact 240ggggacgagg ccgattatta ctgcggaaca tgggatcaca gccggagtgg tgcggtgttc 300ggcggaggga ccaaactgac cgtctta 327 124 327 DNA Homo sapiens anti-Rh(D)chain L03 124 gccgagctca ctcagccacc ctcagcgtct gggacccccg ggcagagggtcaccatctct 60 tgttctggca gtagctccaa catcggaaat aatcatgtaa gctggtaccagcaactccca 120 ggaatggccc ccaaactcct catctattct aatggtcagc ggccctcaggggtccctgac 180 cgattctctg gctccaagtc tggcacctca gcctccctgg ccatcagcggcctccagtct 240 gaggatgagg ctgattatta ttgtgcagca tggcatgaca gcctctatggtccggtgttc 300 ggcggaggga ccaagctgac cgtcctc 327 125 327 DNA Homosapiens anti-Rh(D) chain L04 125 gccgagctca ctcagccacc ctcagcgtctgggacccccg ggcagagggt cagcatctct 60 tgttctggaa gcagctccaa catcggaagtaatactgtaa actggtacca gcagctccca 120 ggaacagccc ccaaactcct catctctactaataatcagg ggccctcagg agtccctgac 180 cgattctctg gctccaagtc tggcacctcatcctccctgg ccatcagtgg gctccggtca 240 gaggctgagg atgattatta ctgtgcagcatgggatgaca ccctgaatgg tgtggtattc 300 ggcggaggga ccaaactgac cgtccta 327126 327 DNA Homo sapiens anti-Rh(D) chain L05 126 gccgagctca ctcagccaccctcagcgtct gggactcccg ggctgagggt caccatctct 60 tgttctggaa gcagctccaacatcggaagt aatattgtaa actggtacca gcagctccca 120 ggaacggccc ccaaactcctcatctttagt aataataagc ggccctcagg ggtccctgac 180 cgattctctg gctccaagtctggcacctca gcctccctgg ccatcagtgg gctccagtct 240 gaggatgagg ctgattattactgtgctaca tgggatgaca gcctgaatgg tcgggtgttc 300 ggcggaggga ccaagctgaccgtccta 327 127 327 DNA Homo sapiens anti-Rh(D) chain M01 127 gccgagctcactcagccacc ctcagcgtct gggacccccg ggcagcgggt caccatctct 60 tgttctgggagcaacttcaa catcggaagt aattatgtat tctggtacca gcatgttcca 120 ggaacggccccaaaactcct catctataat aataatcaac gcccctctgg ggtccctgac 180 cgactctctggctccaagtc tggcgcctca gcctccctgg ccatcaatgg gctccggtcc 240 gatgatgaggctgattatta ctgtacagga tgggatgacc gcctgagtgg cctgattttc 300 ggcggagggccaaaagtgac cgtccta 327 128 327 DNA Homo sapiens anti-Rh(D) chain M02 128gccgagctca cgcagccgcc ctcagcgtct gggacccccg ggcagagggt caccatctct 60tgttctggaa gcagctccaa catcggaagt aattatgtat attggtacca gcagctccca 120ggaacggccc ccaaactcct catctatagg aataatcagc ggccctcagg ggtccctgac 180cgattctctg gctccaagtc tggcacctca gcctccctgg ccatcagtgg gctccggtcc 240gaggatgagg ctgattatta ctgtgcagca tgggatgaca gcctgagtgg ttgggtgttc 300ggcggaggga ccaagctgac cgtccta 327 129 327 DNA Homo sapiens anti-Rh(D)chain M03 129 gccgagctca ctcagccacc ctcagcgtct gggacccccg ggcagagggtcaccatctct 60 tgttctggaa gcagctccaa catcggaagt aattatgtat actggtaccagcagctccca 120 ggaacggccc ccaaactcct catctatagg aataatcagc ggccctcaggggtccctgac 180 cgattctctg gctccaagtc tggcacctca gcctccctgg ccatcagtgggctccggtcc 240 gaggctgagg ctgattatta ctgtgcggca tgggatgaca gcctgagtgccgtggtattc 300 ggcggaggga ccaaactgac cgtccta 327 130 327 DNA Homosapiens anti-Rh(D) chain N01 130 gccgagctca cgcagccgcc ctcagtgtctgcggccccag gacagaaggt caccatctcc 60 tgctctggaa gcagctccaa cattgacagtaactatgtat cctggtacca gcagctccca 120 ggaacagccc ccaaactcct catttttgacaattataggc gaccctcagg gattcctgac 180 cgattctcag gctccaagtc tggcacgtcagccaccctgg gcatcaccgg actccagact 240 ggggacgagg ccgattatta ctgtgcaacatgggatgaca gcctgaatgg tcgggtgttc 300 ggcggaggga ccaagctgac cgtccta 327131 342 DNA Homo sapiens anti-Rh(D) chain N02 131 gccgagctca cgcagccgccctcagtgtct gcggccccag gacagaaggt caccatctcc 60 tgctctggaa gcagctccaacattgggaat aattatgtgt cctggtacca gcaactccca 120 ggaacagccc ccaaactcctcatttatgac aataataagc gaccctcagg gattcctgac 180 cgattctctg gctccaagtctggcacgtca gccaccctgg gcatcaccgg actccagact 240 ggggacgagg ccgattattactgcggaaca tgggatagca gcctgagtgc tggccgcgtt 300 cggcggatgt tcggcggagggaccaagttg accgtcctgg gt 342 132 330 DNA Homo sapiens anti-Rh(D) chainO01 132 gccgagctca cgcagccgcc ctcagtgtct ggggccccag ggcagagggtcaccatctcc 60 tgcactggga gcagctccaa catcggggca ccttatggtg tacactggtaccagcagttt 120 ccaggaacag cccccaaact cgtcatctac aatgacaaca atcggccctcaggggtccct 180 gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcactgggctccag 240 gctgaggatg aggctgatta ttactgccag tcctatgaca gcagcctgagtggaagggtg 300 ttcggcggag ggaccaagct gaccgtccta 330 133 336 DNA Homosapiens anti-Rh(D) chain O02 133 gccgagctca cgcagccgcc ctcagtgtctggggccccag ggcagacggt caccatctcc 60 tgcactggga gcagctccag catcggggcacgttatgatg tacactggta ccaacacctt 120 ccaggaacag cccccaaact cctcatctatggtaaccaca atcggccctc aggggtccct 180 gaccgattct ctggctccaa gtctggcacctcagcctccc tggccatcac tgggctccag 240 gctgaggatg aggctgaata ttattgccagtcctatgaca acagcctgag tggttcgtct 300 gtctttttcg gcggagggac caagctgaccgtccta 336 134 330 DNA Homo sapiens anti-Rh(D) chain O03 134 gccgagctcacgcagccgcc ctctggggcc ccaggccaga cggtcaccat ctcctgcact 60 gggagcagctccaacatcgg ggcaggttat gatgtacact ggtaccagca gcttccagga 120 acagcccccaaactcctcat ctatggtaac agcaatcggc cctcaggggt ccctgaccga 180 ttctctggctccaagtctgg cacctcagcc tccctggcca tcactgggct ccaggctgag 240 gatgaggctgattattactg ccagtcctat gacagcagcc tgagtggtcc ctatgtggta 300 ttcggcggagggaccaagct gaccgtccta 330 135 324 DNA Homo sapiens anti-Rh(D) chain P01135 gccgagctca ctcagccacc ctcggtgtca gtggccccaa gacagacggc caggattacc 60tgtggggggg acaaaatcgg aagtaacact gtgcattggt accggcagat gtcaggccag 120gcccctgttc tggtcatcta tgaagacaaa aaacgacccc ccgggatccc tgagagattc 180tctggttcca cctcagggac aacggccacc ttgagtatca gtggggccca ggttgaggat 240gaagctgact actactgtta ttcaagagac aacagtggtg atcagagaag ggtgttcggc 300gcagggacca agctgaccgt ccta 324 136 330 DNA Homo sapiens anti-Rh(D) chainQ01 136 gccgagctca ctcagccacc ctccgccact gcctccctgg gaggctcggtcaaactcacc 60 tgcattctgc agagtggcca cagaaattac gccgtcgctt ggcatcaccaagaagcaggg 120 aagggcccgc gatttttgat gacggttacc aatgatggca ggcacatcaagggggacggg 180 atccctgatc gcttctcagg ctccgcctct ggggctgaac gctacctctccatctccggc 240 ctccagtctg aggatgaggg tgactactac tgtcagacct ggggcactggcatgcatgtg 300 ttcggcggag ggaccaaact gaccgtccta 330 137 324 DNA Homosapiens anti-Rh(D) chain R01 137 gccgagctca ctcagcctcc ctccgcgtccgggtctcctg gacagtcagt caccatctcc 60 tgcactggag ccagcagtga cgttggtgcttataagcacg tctcctggta ccaacaacac 120 ccaggcaaag cccccaaact cctgactcatgagggcacta agcggccctc aggggtccct 180 gatcgcttct ctggctccaa gtctggcaacacggcctccc tgaccgtctc tgggctccag 240 gctgaggatg aggctgatta ttactgcagctcatttgcag gtaattccgt gatattcggc 300 ggagggacca agctgaccgt ccta 324 138312 DNA Homo sapiens anti-Rh(D) chain S01 138 gccgagctca ctcagcctccctccgtgtct gggtctcctg gacagtcgat caccatctcc 60 tgcagtgatg ttgggaattataaccttgtc tcctggtacc aacagtaccc aggcaaggcc 120 cccaaactca taatttatgagggcagtaag cggccctcag gggtttctag tcgcttctct 180 ggctccaggt ctggcaacacggcctccctg acaatctctg ggctccaggc tgaggacgag 240 gctgattatc actgctgctcatatgcaatt agtagcagga ttttcggcgg agggaccaag 300 ctgaccgtcc ta 312 139127 PRT Homo sapiens anti-Rh(D) antibody clone SH10 139 Glu Val Gln LeuLeu Glu Glu Ser Gly Gly Gly Val Val Gln Pro Gly 1 5 10 15 Arg Ser LeuArg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg 20 25 30 Asn Gly MetHis Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Ala PheIle Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser 50 55 60 Val Lys GlyArg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70 75 80 Tyr LeuGln Met Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr Tyr 85 90 95 Cys AlaArg Glu Glu Ala Leu Phe Arg Gly Leu Thr Arg Trp Ser Tyr 100 105 110 GlyMet Asp Val Trp Gly Gln Gly Thr Thr Val Ser Val Ser Ser 115 120 125 140125 PRT Homo sapiens anti-Rh(D) antibody clone SH16 140 Glu Val Gln LeuLeu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu ArgLeu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Gly Met HisTrp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Val 35 40 45 Ala Leu IleTrp Tyr Asp Gly Gly Asn Lys Glu Tyr Ala Asp Ser Val 50 55 60 Lys Gly ArgPhe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu GlnVal Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala ArgAsp Gln Arg Ala Ala Ala Gly Ile Phe Tyr Tyr Ser Arg Met 100 105 110 AspVal Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 141 117 PRTHomo sapiens anti-Rh(D) antibody clone SH17 141 Glu Val Gln Leu Leu GluSer Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu SerCys Gly Ala Ser Gly Ile Pro Phe Val Ser Ser 20 25 30 Trp Met Ala Trp ValArg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Asn Ile Lys GlnAsp Gly Ser Lys Lys Asn Tyr Val Asp Ser Val 50 55 60 Glu Gly Arg Phe ThrIle Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met AspSer Leu Arg Ala Glu Asp Thr Arg Ile Tyr Tyr Cys 85 90 95 Ala Arg Asp SerLeu Thr Cys Phe Asp Tyr Trp Gly Gln Gly Ala Leu 100 105 110 Val Thr ValSer Ser 115 142 128 PRT Homo sapiens anti-Rh(D) antibody clone SH18 142Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 1015 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr 20 2530 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 4045 Ala Ala Thr Ala Tyr Asp Gly Lys Asn Lys Tyr Tyr Ala Asp Ser Val 50 5560 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Met Asn Thr Leu Phe 65 7075 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Phe Tyr Cys 8590 95 Ala Arg Gly Gly Phe Tyr Tyr Asp Ser Ser Gly Tyr Tyr Gly Leu Arg100 105 110 His Tyr Phe Asp Ser Trp Gly Gln Gly Thr Leu Val Thr Val SerSer 115 120 125 143 129 PRT Homo sapiens anti-Rh(D) antibody clone SH20143 Glu Val Gln Leu Leu Glu Glu Ser Gly Gly Gly Val Val Gln Pro Gly 1 510 15 Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser 2025 30 Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 3540 45 Val Ala Val Ile Ser Tyr Asp Gly Ser Thr Ile Tyr Tyr Ala Asp Ser 5055 60 Val Lys Gly Arg Phe Thr Ile Ser Arg Ala Asn Ser Lys Asn Thr Leu 6570 75 80 Phe Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Val Tyr Tyr85 90 95 Cys Thr Arg Gly Gly Phe Tyr Tyr Asp Ser Ser Gly Tyr Tyr Gly Leu100 105 110 Arg His Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr ValSer 115 120 125 Ser 144 126 PRT Homo sapiens anti-Rh(D) antibody cloneSH24 144 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Ser Leu Arg SerTyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu TrpVal 35 40 45 Ala Asp Ile Trp Phe Asp Gly Ser Asn Lys Asp Tyr Ala Asp SerVal 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr LeuTyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val TyrTyr Cys 85 90 95 Ala Arg Asp Trp Arg Val Arg Ala Phe Ser Ser Gly Trp LeuSer Ala 100 105 110 Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val SerSer 115 120 125 145 127 PRT Homo sapiens anti-Rh(D) antibody clone SH25145 Glu Val Gln Leu Leu Glu Glu Ser Gly Gly Gly Val Val Gln Pro Gly 1 510 15 Arg Ser Leu Arg Leu Ala Cys Ala Ala Ser Gly Phe Ser Phe Arg Ser 2025 30 Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp 3540 45 Val Ala Phe Thr Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Val Asp Ser 5055 60 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 6570 75 80 Tyr Leu Glu Met Asn Ser Leu Arg Val Asp Asp Thr Ala Val Tyr Tyr85 90 95 Cys Ala Arg Glu Ala Pro Met Leu Arg Gly Ile Ser Arg Tyr Tyr Tyr100 105 110 Ala Met Asp Val Trp Gly Pro Gly Thr Thr Val Thr Val Ser Ser115 120 125 146 126 PRT Homo sapiens anti-Rh(D) antibody clone SH28,SH50, and SH53 146 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Gly Val GlnPro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe ThrPhe Asn Ser Tyr 20 25 30 Ala Met Tyr Trp Val Arg Gln Pro Pro Gly Lys GlyLeu Glu Trp Val 35 40 45 Ala Ala Ile Trp Tyr Asp Gly Ser Asn Lys Glu TyrAla Asp Phe Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser LysAsn Thr Leu Ser 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Asp Glu Asp ThrAla Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Asn Leu Leu Arg Gly Trp SerArg Tyr Tyr Tyr Gly 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr ValThr Val Ser Ser 115 120 125 147 126 PRT Homo sapiens anti-Rh(D) antibodyclone SH32 147 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln ProGly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Lys Phe Thr LeuTyr Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly LeuGlu Trp Val 35 40 45 Ala Phe Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr GluAsp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys AsnThr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr AlaVal Tyr Tyr Cys 85 90 95 Ala Arg Glu Leu Ser Lys Lys Val Ala Leu Ser ArgTyr Tyr Tyr Tyr 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val ThrVal Ser Ser 115 120 125 148 126 PRT Homo sapiens anti-Rh(D) antibodyclone SH37 148 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln ProGly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Lys Phe Thr LeuTyr Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly LeuGlu Trp Val 35 40 45 Ala Phe Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr GluAsp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys AsnThr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr AlaVal Tyr Tyr Cys 85 90 95 Ala Arg Glu Leu Ser Lys Lys Val Ala Leu Ser ArgTyr Tyr Tyr Tyr 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val ThrVal Ser Ser 115 120 125 149 126 PRT Homo sapiens anti-Rh(D) antibodyclone SH39 149 Glu Val Gln Leu Leu Glu Gln Ser Gly Gly Gly Val Val GlnPro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe ThrPhe Ser Ser 20 25 30 Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys GlyLeu Glu Trp 35 40 45 Val Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Glu TyrAla Asp Ser 50 55 60 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser LysAsn Thr Leu 65 70 75 80 Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp ThrAla Val Tyr Tyr 85 90 95 Cys Ala Arg Glu Glu Val Val Arg Gly Val Ile LeuTrp Ser Arg Lys 100 105 110 Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val ThrVal Ser Ser 115 120 125 150 126 PRT Homo sapiens anti-Rh(D) antibodyclone SH44 150 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Ala Gln ProGly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Ser LeuArg Ser Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly LeuGlu Trp Val 35 40 45 Ala Asp Ile Trp Phe Asp Gly Ser Asn Lys Asp Tyr AlaAsp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys AsnThr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr AlaVal Tyr Tyr Cys 85 90 95 Ala Arg Asp Trp Arg Val Arg Ala Phe Ser Ser GlyTrp Leu Ser Ala 100 105 110 Phe Asp Ile Trp Gly Gln Gly Thr Met Val ThrVal Ser Ser 115 120 125 151 125 PRT Homo sapiens anti-Rh(D) antibodyclone SH47 151 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln ProGly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser PheSer Asn Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly LeuGlu Trp Val 35 40 45 Ala Val Thr Ser Phe Asp Gly Ser Ile Lys Asp Tyr AlaAsp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys AsnThr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr AlaVal Tyr Tyr Cys 85 90 95 Ala Arg Glu Arg Gly Met Ile Val Val Val Arg ArgArg Asn Ala Phe 100 105 110 Asp Ile Trp Gly Gln Gly Thr Met Val Thr ValSer Ser 115 120 125 152 126 PRT Homo sapiens anti-Rh(D) antibody cloneSH54 152 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser ArgAsn 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu TrpVal 35 40 45 Ala Phe Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Ala Asp SerVal 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr LeuTyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Asp Asp Thr Ala Val TyrTyr Cys 85 90 95 Ala Arg Glu Glu Ala Leu Phe Arg Gly Leu Thr Arg Trp SerTyr Gly 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val Ser Val SerSer 115 120 125 153 126 PRT Homo sapiens anti-Rh(D) antibody clone SH56153 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 510 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 2025 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 3540 45 Ala Val Val Tyr Tyr Asp Gly Ser Asn Lys His Tyr Ser Asp Ser Val 5055 60 Lys Gly Arg Phe Thr Ile Phe Arg Asp Asn Ser Lys Asn Thr Leu Tyr 6570 75 80 Leu Gln Met Asp Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys85 90 95 Ala Arg Glu Arg Asn Phe Arg Ser Gly Tyr Ser Arg Tyr Tyr Tyr Gly100 105 110 Met Asp Val Trp Gly Pro Gly Thr Thr Val Thr Val Ser Ser 115120 125 154 107 PRT Homo sapiens anti-Rh(D) antibody clone SH8 154 AlaGlu Leu Thr Gln Ser Pro Ser Ser Leu Ala Ala Ser Val Gly Asp 1 5 10 15Arg Val Thr Ile Thr Cys Arg Ala Asn Gln Thr Ile Arg Thr Ser Leu 20 25 30Asn Trp Tyr Gln Gln Arg Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 40 45Gly Ala Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly Gly 50 55 60Ile Ser Gly Ala Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Tyr Gly Tyr Ser Arg Thr 85 9095 Phe Gly Gln Gly Thr Lys Val Asp Ile Lys Arg 100 105 155 107 PRT Homosapiens anti-Rh(D) antibody clone SH12 155 Ala Glu Leu Thr Gln Ser ProPhe Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr CysArg Ala Ser His Asn Ile Tyr Arg Ser Leu 20 25 30 Asn Trp Phe Gln His LysPro Gly Glu Ala Pro Lys Leu Leu Val Tyr 35 40 45 Ala Ala Ser Ser Leu GlnArg Gly Val Pro Thr Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp PheThr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Ser Ala Thr TyrPhe Cys Gln Gln Ser Val Thr Phe Pro Tyr Thr 85 90 95 Phe Gly Gln Gly ThrLys Leu Glu Ile Arg Arg 100 105 156 107 PRT Homo sapiens anti-Rh(D)antibody clone SH13 156 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln SerIle Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Arg Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile SerSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln SerTyr Ser Thr Pro Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile LysArg 100 105 157 107 PRT Homo sapiens anti-Rh(D) antibody clone SH14 157Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 1015 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Arg Arg Ser Leu 20 2530 Asn Trp Tyr Gln His Lys Pro Gly Arg Ala Pro Arg Leu Leu Ile Tyr 35 4045 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Arg Gly Ser 50 5560 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro Ala 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn Thr Pro Trp Thr 8590 95 Phe Gly His Gly Thr Lys Val Glu Ile Lys Arg 100 105 158 107 PRTHomo sapiens anti-Rh(D) antibody clone SH16 158 Ala Glu Leu Thr Gln SerPro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile ThrCys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser LeuGln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr AspPhe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro Thr 85 90 95 Phe Gly Gly GlyThr Lys Val Glu Ile Lys Arg 100 105 159 106 PRT Homo sapiens anti-Rh(D)antibody clone SH18 159 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln SerIle Ser Ile Ala Leu 20 25 30 Asn Trp Tyr Gln Gln Arg Pro Gly Lys Ala ProLys Leu Leu Met Tyr 35 40 45 Ala Thr Ser Thr Leu Gln Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile SerSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln TyrTyr Asn Lys Pro Thr Phe 85 90 95 Gly Pro Gly Thr Lys Val Asp Ile Lys Arg100 105 160 107 PRT Homo sapiens anti-Rh(D) antibody clone SH20 160 AlaGlu Leu Thr Gln Ser Pro Phe Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Arg Ser Leu 20 25 30Asn Trp Tyr Gln His Lys Pro Gly Glu Ala Pro Lys Leu Leu Ile Tyr 35 40 45Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Pro Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 7580 Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ser Val Arg Ile Pro Tyr Ser 85 9095 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 161 108 PRT Homosapiens anti-Rh(D) antibody clone SH21 161 Ala Glu Leu Thr Gln Ser ProSer Phe Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr CysArg Ala Ser Gln Gly Ile Arg Ser Tyr Leu 20 25 30 Ala Trp Tyr Gln Gln LysPro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Thr Leu GlnSer Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu PheThr Leu Thr Ile Ala Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr TyrTyr Cys Gln Gln Leu Asn Asn Tyr Pro Pro Phe 85 90 95 Thr Phe Gly Pro GlyThr Lys Val Asp Ile Lys Arg 100 105 162 107 PRT Homo sapiens anti-Rh(D)antibody clone SH24 162 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln SerIle Ser Thr Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Arg Pro Gly Lys Ala ProAsn Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Thr Leu Gln Arg Gly Val Pro SerArg Phe Thr Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile SerSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln SerTyr Thr Thr Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Met Glu Ile ArgArg 100 105 163 108 PRT Homo sapiens anti-Rh(D) antibody clone SH26 163Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 1015 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 2530 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 4045 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 5560 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Phe Arg Arg Tyr 8590 95 Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 164 107PRT Homo sapiens anti-Rh(D) antibody clone SH28 164 Ala Glu Leu Thr GlnSer Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr IleThr Cys Arg Ala Asp Gln Asn Ile Arg Arg Ser Leu 20 25 30 Asn Trp Phe GlnGln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser SerLeu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly ThrAsp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe AlaThr Tyr Tyr Cys Gln Gln Ser Ser Ser Thr Pro Trp Thr 85 90 95 Phe Gly ArgGly Thr Lys Val Glu Ile Lys Arg 100 105 165 106 PRT Homo sapiensanti-Rh(D) antibody clone SH30 165 Ala Glu Leu Thr Gln Ser Pro Ser SerLeu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg AlaSer Gln Ser Ile Arg Arg Ser Leu 20 25 30 Asn Trp Tyr Gln Gln Ser Pro GlyLys Thr Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser GlyVal Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr LeuThr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr CysGln Gln Ser Tyr Ser Thr Leu Thr Phe 85 90 95 Gly Gly Gly Thr Lys Val GluIle Lys Arg 100 105 166 108 PRT Homo sapiens anti-Rh(D) antibody cloneSH32 166 Ala Glu Leu Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr1 5 10 15 Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser ArgTyr 20 25 30 Phe Pro Asn Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg AlaLeu 35 40 45 Ile Tyr Gly Ser Asn Asn Lys His Ser Trp Thr Pro Ala Arg PheSer 50 55 60 Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly ValGln 65 70 75 80 Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Leu Leu Phe Tyr AlaGly Ala 85 90 95 Trp Ala Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105167 108 PRT Homo sapiens anti-Rh(D) antibody clone SH34 167 Ala Glu LeuThr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg ValThr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn TrpTyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala AlaSer Gly Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly SerGly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 AspPhe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro Tyr 85 90 95 ThrPhe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 168 107 PRT Homosapiens anti-Rh(D) antibody clone SH36 168 Ala Glu Leu Thr Gln Ser ProSer Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr CysArg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln LysPro Gly Lys Ser Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu GlnSer Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp PheThr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr TyrTyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro Ala 85 90 95 Phe Gly Pro Gly ThrLys Val Asp Ile Lys Arg 100 105 169 107 PRT Homo sapiens anti-Rh(D)antibody clone SH39 169 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln ThrIle Gly Arg Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Arg Pro Gly Lys Ala ProLys Leu Leu Val Tyr 35 40 45 Ala Val Ser Ser Leu Gln Ser Gly Ala Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr His Phe Thr Leu Thr Ile ThrSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Phe Cys Gln Gln SerTyr Ser Ser Pro Phe Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile LysArg 100 105 170 107 PRT Homo sapiens anti-Rh(D) antibody clone SH41 170Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 1015 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Arg Arg Ser Leu 20 2530 Asn Trp Tyr Gln His Lys Pro Gly Arg Ala Pro Arg Leu Leu Ile Tyr 35 4045 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Arg Gly Ser 50 5560 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro Ala 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn Thr Pro Trp Thr 8590 95 Phe Gly His Gly Thr Lys Val Glu Ile Lys Arg 100 105 171 106 PRTHomo sapiens anti-Rh(D) antibody clone SH44 171 Ala Glu Leu Thr Gln SerPro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Ile Ile ThrCys Arg Ala Ser Gln Thr Ile Pro Arg Phe Leu 20 25 30 Asn Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Val Leu Leu Ile His 35 40 45 Ser Ile Ser Ser LeuGln Ser Gly Val Pro Ser Arg Phe Ser Ala Ser 50 55 60 Gly Ser Gly Thr GluPhe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Gln Gln Ser Tyr Ser Asn Leu Ser Phe 85 90 95 Gly Pro Gly ThrThr Val Asp Ile Arg Arg 100 105 172 107 PRT Homo sapiens anti-Rh(D)antibody clone SH46 172 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln TyrIle Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProAsn Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile SerSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln ThrTyr Ser Ser Pro Ser Thr 85 90 95 Phe Gly Pro Gly Thr Lys Val Asp Ile LysArg 100 105 173 107 PRT Homo sapiens anti-Rh(D) antibody clone SH47 173Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 1015 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Asn Tyr Leu 20 2530 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 4045 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 5560 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Tyr Pro Arg Thr 8590 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Arg Arg 100 105 174 107 PRTHomo sapiens anti-Rh(D) antibody clone SH48 174 Ala Glu Leu Thr Gln SerPro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile ThrCys Arg Ala Ser Gln Tyr Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser LeuGln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr AspPhe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Gln Gln Thr Tyr Ser Ser Pro Ser Thr 85 90 95 Phe Gly Pro GlyThr Lys Val Asp Ile Lys Arg 100 105 175 107 PRT Homo sapiens anti-Rh(D)antibody clone SH49 175 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser AlaSer Val Gly Asp 1 5 10 15 Arg Val Thr Val Thr Cys Arg Ala Ser Gln SerIle Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile SerSer Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln SerTyr Ser Thr Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile LysArg 100 105 176 107 PRT Homo sapiens anti-Rh(D) antibody clone SH50 176Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 1015 Arg Val Thr Val Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 2530 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 4045 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 5560 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Trp Thr 8590 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 177 108 PRTHomo sapiens anti-Rh(D) antibody clone SH51 177 Ala Glu Leu Thr Gln SerPro Ser Phe Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile ThrCys Arg Ala Ser Gln Gly Ile Arg Ser Tyr Leu 20 25 30 Ala Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Thr LeuGln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr GluPhe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Gln Gln Leu Asn Asn Tyr Pro Pro Phe 85 90 95 Thr Phe Gly ProGly Thr Lys Val Asp Ile Lys Arg 100 105 178 108 PRT Homo sapiensanti-Rh(D) antibody clone SH52 178 Ala Glu Leu Thr Gln Ser Pro Gly ThrLeu Ser Leu Ser Pro Gly Glu 1 5 10 15 Arg Ala Thr Leu Ser Cys Arg AlaSer Gln Ser Ile Ser Ser Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys ProGly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Ser Arg Ala ThrGly Ile Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe ThrLeu Thr Ile Ser Arg Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr TyrCys Gln Gln Tyr Gly Ser Ser Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr LysVal Glu Ile Lys Arg 100 105 179 107 PRT Homo sapiens anti-Rh(D) antibodyclone SH54 179 Ala Glu Leu Thr Gln Ser Pro Ser Ser Met Ser Ala Ser ValGly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile GlyThr Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys LeuLeu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg PheSer Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser LeuGln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr SerThr Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100105 180 109 PRT Homo sapiens anti-Rh(D) antibody clone SH55 180 Ala GluLeu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg 1 5 10 15 ValThr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Lys Tyr 20 25 30 ValTyr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 35 40 45 TyrSer Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Ala 50 55 60 PheLys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln Ala 65 70 75 80Glu Asp Glu Ala Asn Tyr Tyr Cys Gln Ser Tyr Asp Ser Gly Leu Ser 85 90 95Gly Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 181 108 PRTHomo sapiens anti-Rh(D) antibody clone SH56 181 Ala Glu Leu Thr Gln SerPro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile ThrCys Arg Ala Ser Gln Ser Ile Ser Arg Tyr Leu 20 25 30 Asn Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser LeuGln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr AspPhe Ala Leu Thr Ile Ser Ser Leu Leu Pro Glu 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Gln Gln Gly Tyr Ser Thr Pro Pro Tyr 85 90 95 Ser Phe Gly GlnGly Thr Lys Leu Glu Ile Lys Arg 100 105 182 381 DNA Homo sapiensanti-Rh(D) antibody clone SH10 182 gaggtgcagc tgctcgagga gtctgggggaggcgtggtcc agcctgggag gtccctgaga 60 ctctcctgtg cagcgtctgg gttcaccttcagtaggaatg gcatgcactg ggtccgccag 120 gctcctggca aggggctgga gtgggtggcatttatatggt ttgatggaag taataaatac 180 tatgcagact ccgtgaaggg ccgattcaccatctccagag acaattccaa gaacacgctg 240 tatctgcaaa tgaacagcct gagagccgacgacacggctg tgtattactg tgcgagagag 300 gaggctctgt ttcggggact tactcggtggtcctacggca tggacgtctg gggccaaggg 360 accacggtca gcgtctcctc a 381 183 375DNA Homo sapiens anti-Rh(D) antibody clone SH16 183 gaggtgcagctgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcagcgtctgggtt caccttcagt agctatggca tgcactgggt ccgccaggct 120 ccaggcagggggctggagtg ggtggctctt atatggtacg atggaggtaa caaagagtat 180 gcagactccgtgaagggccg cttcagcatc tccagagaca actccaagaa cactctgtat 240 ctgcaagtgaacagcctgag agccgacgac acggctgtct attactgtgc gagagaccag 300 agagcagcagcgggtatctt ttattattcc cgtatggacg tctggggcca agggaccacg 360 gtcaccgtctcctca 375 184 351 DNA Homo sapiens anti-Rh(D) antibody clone SH17 184gaggtgcagc tgctcgagtc tgggggaggc ttggtccagc cgggggggtc cctgagactc 60tcctgtggtg cctctggaat cccctttgtt tcctcttgga tggcctgggt ccgccaggcc 120ccagggaagg ggctggagtg ggtggccaac ataaaacaag atggaagtaa gaaaaactat 180gtggactctg tggagggccg attcaccatc tccagagaca acgcgaagaa ctcactttat 240ctgcaaatgg acagcctgag agccgaggac acgcggatat attactgtgc gcgagattca 300cttacttgtt ttgactactg gggccaggga gccctggtca ccgtctcctc a 351 185 384 DNAHomo sapiens anti-Rh(D) antibody clone SH18 185 gaggtgcagc tgctcgagtctgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cctctggattcaccttcagg agctatgcta tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtgggtggcagct acagcatatg atggaaaaaa taaatactac 180 gcagactccg tgaagggccgattcaccatc tccagagaca attccatgaa cacgctgttt 240 ctgcaaatga acagcctgagagctgaggac acggctgtgt tttactgtgc gagaggcgga 300 ttttactatg atagtagtggttattacggc ttgaggcact actttgactc ctggggccag 360 ggaaccctgg tcaccgtctcctca 384 186 387 DNA Homo sapiens anti-Rh(D) antibody clone SH20 186gaggtgcagc tgctcgagga gtctggggga ggcgtggtcc agcctgggag gtccctgaga 60ctctcctgtg cagcctctgg attcaccttc agaagttatg ctatgcactg ggtccgccag 120gctccaggca aggggctgga gtgggtggcg gttatatcat atgatggaag tactatatac 180tacgcagact ccgtgaaggg ccgattcacc atctccagag ccaattccaa gaacacgctg 240tttctgcaaa tgaacagcct cagaactgag gacacggctg tatattactg tacgagaggg 300gggttttact atgacagtag tggttattac gggttgaggc actactttga ctactggggc 360cagggaaccc tggtcaccgt ctcttca 387 187 378 DNA Homo sapiens anti-Rh(D)antibody clone SH24 187 gaggtgcagc tgctcgagtc ggggggaggc gtggcccagcctgggaggtc cctgagactc 60 tcctgtgtag cgtctggatt cagcctcagg agctatggcatgcactgggt ccgccaggct 120 cctggcaagg ggctggagtg ggtggcagat atatggtttgatggaagtaa taaagattat 180 gcagactccg tgaagggccg attcaccatc tccagagacaattccaagaa cacgttgtat 240 cttcaaatga acagcctgag agccgaggac acggctgtgtattattgtgc gagagattgg 300 agggtgcggg cctttagtag tggctggtta agtgcttttgatatctgggg ccaagggaca 360 atggtcaccg tctcttca 378 188 381 DNA Homosapiens anti-Rh(D) antibody clone SH25 188 gaggtgcagc tgctcgaggagtctggggga ggcgtggtcc agcctgggag gtccctgaga 60 ctcgcctgtg cagcgtctggattcagcttc aggagctatg gcatgcactg ggtccgccag 120 gctccaggca gggggctggagtgggtggca tttacatggt ttgatggaag caataaatat 180 tatgtagact ccgtgaagggccgattcacc atctccagag acaattccaa gaacacgctg 240 tatctggaaa tgaacagcctgagagtcgat gacacggctg tatattactg tgcgagagag 300 gcgcctatgc ttcgcggaattagcagatac tactacgcga tggacgtctg gggcccaggg 360 accacggtca ccgtctcctc a381 189 378 DNA Homo sapiens anti-Rh(D) antibody clone SH28, SH50, andSH53 189 gaggtgcagc tgctcgagtc tgggggaggc ggggtccagc ctgggaggtccctgcgactc 60 tcctgtgcgg cgtctggatt caccttcaat agttatgcca tgtactgggtccgccagcct 120 ccaggcaagg ggctggagtg ggtggcagct atatggtatg atggaagtaataaagaatat 180 gcagattttg tgaagggccg cttcaccatc tccagagaca attccaagaacacgctgtct 240 ctgcaaatga acagcctgag agacgaggac acggctgtgt attactgtgcgagagaggcg 300 aatctcctcc gtggctggtc tcgatactac tacggtatgg acgtctggggccaagggacc 360 acggtcaccg tctcctca 378 190 378 DNA Homo sapiensanti-Rh(D) antibody clone SH32 190 gaggtgcagc tgctcgagtc ggggggaggcgtggtccagc ctgggaggtc cctgagactc 60 tcctgtgaag cgtctaaatt caccctctacaattatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcatttatatggtttg atggaagtaa taaatactat 180 gaagactccg tgaagggccg attcaccgtctccagagaca attccaagaa cacgctgtat 240 ctgcaaatga acagcctgag agccgaggacacggctgtgt attactgtgc gagagaacta 300 tctaagaagg tggcactttc taggtattactactatatgg acgtctgggg ccaggggacc 360 acggtcactg tctcgtca 378 191 378 DNAHomo sapiens anti-Rh(D) antibody clone SH37 191 gaggtgcagc tgctcgaggagtctggggga ggcgtggtcc agcctgggag gtccctgaga 60 ctctcctgtg cagtgtctggattcacccta actaattatg gcatgcactg ggtccgccag 120 gctccaggca aggggctggagtgggtggca catgtctggt atgatggaag taaaacagaa 180 tacgcagact ccgtcaagggccgattcgcc gtctccagag acaaatccaa gaacacactg 240 tttctgcaaa tgaacagcctgacagccgag gacacggcta tttattactg tgcgagagag 300 aggagagaga aagtctatatattgttctac tcgtggctcg accgctgggg ccagggaacc 360 ctggtcaccg tctcctca 378192 378 DNA Homo sapiens anti-Rh(D) antibody clone SH39 192 gaggtgcagctgctcgagca gtctggggga ggcgtggtcc agcctgggag gtccctgaga 60 ctctcctgtgcagcgtctgg attcaccttc agtagctatg gcatgcactg ggtccgccag 120 gctccaggcaagggactgga gtgggtggca gttatatggt ttgatggaag taataaggaa 180 tatgcagactccgtgaaggg ccgattcacc atctccagag acaattccaa gaacacgctg 240 tatctacaaatgaacagcct gagagccgag gacacggctg tgtattactg tgcgagagaa 300 gaagtggttcggggagttat cttatggtct cggaagtttg actactgggg ccagggaacc 360 ctggtcaccgtctcctca 378 193 378 DNA Homo sapiens anti-Rh(D) antibody clone SH44 193gaggtgcagc tgctcgagtc ggggggaggc gtggcccagc ctgggaggtc cctgagactc 60tcctgtgtag cgtctggatt cagcctcagg agctatggca tgcactgggt ccgccaggct 120cctggcaagg ggctggagtg ggtggcagat atatggtttg atggaagtaa taaagattat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgttgtat 240cttcaaatga acagcctgag agccgaggat acggctgtgt attattgtgc gagagattgg 300agggtgcggg cctttagtag tggctggtta agtgcttttg atatctgggg ccaagggaca 360atggtcaccg tctcttca 378 194 375 DNA Homo sapiens anti-Rh(D) antibodyclone SH47 194 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtccctgcgactc 60 tcttgtgcag cctctggatt cagcttcagt aactatgcta tgcactgggtccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagtt acatcatttg atggaagcattaaagactac 180 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaacacactatat 240 ctgcaaatga acagcctgag agatgaggac acggctgtat attactgtgcgagagagcgg 300 gggatgatag tcgtggtccg tcgcagaaat gcttttgata tttggggccaagggacaatg 360 gtcaccgtct cttca 375 195 378 DNA Homo sapiens anti-Rh(D)antibody clone SH54 195 gaggtgcagc tgctcgagtc ggggggaggc gtggtccagcctgggaggtc cctgagactc 60 tcctgtgcag cgtctgggtt caccttcagt aggaatggcatgcactgggt ccgccaggct 120 cctggcaagg ggctggagtg ggtggcattt atatggtttgatggaagtaa taaatactat 180 gcagactccg tgaagggccg attcaccatc tccagagacaattccaagaa cacgctgtat 240 ctgcaaatga acagcctgag agccgacgac acggctgtgtattactgtgc gagagaggag 300 gctctgtttc ggggacttac tcggtggtcc tacggtatggacgtctgggg ccaagggacc 360 acggtcagcg tctcctca 378 196 378 DNA Homosapiens anti-Rh(D) antibody clone SH56 196 gaggtgcagc tgctcgagtcggggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cgtctggattcaccttcagt agctatggca tgcactgggt ccggcaggct 120 ccaggcaagg ggctggagtgggtggcagtt gtctactatg atggaagtaa caaacactat 180 tcagactccg tgaagggccgattcaccatc ttcagagaca actccaagaa cacgctgtat 240 ctacaaatgg acagcctgagagccgaggac acggctgtgt attactgtgc gagagaaaga 300 aattttcgga gtggttattcccgctactac tacggtatgg acgtctgggg cccagggacc 360 acggtcaccg tctcctca 378197 321 DNA Homo sapiens anti-Rh(D) antibody clone SH8 197 gccgagctcacccagtctcc atcctccctg gctgcgtctg tcggagacag agtcaccatc 60 acttgccgggcaaatcagac catcagaacc tctttaaatt ggtatcaaca aagacctggg 120 aaagcccctaacctcctgat ctatggtgca tccaggttgc atagtggggt cccatcaagg 180 tttagtggcggtatttctgg ggcagacttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaacttactactg tcagcagact tacggttatt ctcgaacgtt cggccaaggg 300 accaaggtggatatcaaacg a 321 198 321 DNA Homo sapiens anti-Rh(D) antibody clone SH12198 gccgagctca cccagtctcc attctccctg tctgcatctg taggagacag agtcaccata 60acttgccggg caagtcacaa catttacagg tctttaaatt ggtttcagca taaaccaggg 120gaagccccta agctcctggt ctatgctgca tccagtctgc agcgtggggt cccaaccagg 180ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct tcaacctgaa 240gactctgcga cttacttctg tcaacagagt gtcacattcc cctacacttt tggccagggg 300accaagctgg agatcagacg a 321 199 321 DNA Homo sapiens anti-Rh(D) antibodyclone SH13 199 gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacagagtcaccatc 60 acttgccggg caagtcagag cattagcagc tatttaaatt ggtatcagcagaaaccaggg 120 aaagccccta agctcctgat ctatgctgca tccagtttgc gaagtggggtcccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtctgcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tacagtaccc cctacacttttggccagggg 300 accaagctgg agatcaaacg a 321 200 321 DNA Homo sapiensanti-Rh(D) antibody clone SH14 200 gccgagctca cccagtctcc atcctccctgtctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagaa cattaggaggtctttaaatt ggtatcaaca caaaccaggg 120 agagccccta gactcctgat ctatgctgcatccactttgc aaagtggggt cccatcaagg 180 ttcaggggca gtggatctgg gacagatttcactctcacca tcaacagtct gcaacctgca 240 gattttgcaa cttactactg tcagcagagttccaataccc cgtggacgtt cggccatggg 300 accaaggtgg aaatcaaacg a 321 201 321DNA Homo sapiens anti-Rh(D) antibody clone SH16 201 gccgagctcacccagtctcc atcctccctg tctgcctctg taggagacag agtcaccatc 60 acttgccgggcaagtcagag cattagcagc tatttaaatt ggtatcaaca gaaaccaggg 120 aaagcccctaagctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggcagtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaacttactactg tcaacagagt tacagtaccc ctccaacttt cggcggaggg 300 accaaggtggagatcaaacg a 321 202 318 DNA Homo sapiens anti-Rh(D) antibody clone SH18202 gccgagctca cccagtctcc atcctccctc tctgcatctg taggagacag agtcaccatc 60acttgccggg caagtcagag tattagcatc gctttaaatt ggtatcagca gagaccaggg 120aaagccccta agctcctgat gtatgctaca tccactttgc aaagtggggt cccatcaagg 180ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240gattttgcaa cttactactg tcaacaatat tacaataaac ctactttcgg ccctgggacc 300aaggtggata tcaaacga 318 203 321 DNA Homo sapiens anti-Rh(D) antibodyclone SH20 203 gccgagctca cccagtctcc attctccctg tctgcatctg tcggagacagagtcaccata 60 acttgccggg caagtcagag cattagcagg tctttaaatt ggtatcaacataaaccaggg 120 gaagccccta agctcctgat ctatgctgca tccagtctgc agcgtggggtcccacccagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtctgcaacctgaa 240 gactttgcga cttacttctg tcaacagagt gtcagaatcc cgtacagttttggccagggg 300 accaagctgg agatcaaacg a 321 204 324 DNA Homo sapiensanti-Rh(D) antibody clone SH21 204 gccgagctca cccagtctcc atccttcctgtctgcatctg taggagacag agtcaccatc 60 acttgccggg ccagtcaggg cattaggagttatttagcct ggtatcagca aaaaccaggg 120 aaagccccta agctcctaat ctatgctgcatccactttgc aaagtggggt cccatcaagg 180 ttcagcggca gtggatctgg gacagaattcactctcacaa tcgccagcct gcagcctgat 240 gattttgcaa cttattactg tcaacagcttaataattacc cccctttcac tttcggccct 300 gggaccaaag tggatatcaa acga 324 205321 DNA Homo sapiens anti-Rh(D) antibody clone SH24 205 gccgagctcacccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc 60 acttgccgggcaagtcagag cattagcacc tatttaaatt ggtatcagca gagaccaggg 120 aaagcccctaacctcctgat ctatgctgca tccactttgc aaaggggggt cccatcaagg 180 ttcactggcagtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaacttactactg tcaacagagt tacactaccc tgtggacgtt cggccaaggg 300 accaagatggaaatcagacg a 321 206 324 DNA Homo sapiens anti-Rh(D) antibody clone SH26206 gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc 60acttgccggg caagtcagag cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240gattttgcaa cttactactg tcaacagagt tacagtttcc gaaggtacag ttttggccag 300gggaccaagc tggagatcaa acga 324 207 321 DNA Homo sapiens anti-Rh(D)antibody clone SH28 207 gccgagctca cccagtctcc atcctccctg tctgcatctgtaggagacag agtcaccatc 60 acttgccggg cagatcagaa cattaggagg tctttaaattggtttcagca gaaaccaggg 120 aaagccccta agctcctgat ctatgctgca tccagtttgcaaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcaccatcagcagtct gcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tccagtaccccgtggacgtt cggccgaggg 300 accaaggtgg aaatcaaacg a 321 208 318 DNA Homosapiens anti-Rh(D) antibody clone SH30 208 gccgagctca cccagtctccatcctccctg tctgcatctg ttggagacag agtcaccatc 60 acttgccggg caagtcagagcattcggagg tctttaaatt ggtatcagca gagtccaggg 120 aaaaccccta agctcctgatctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgggacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactgtcaacagagt tacagtaccc tcactttcgg cggagggacc 300 aaggtggaga tcaaacga 318209 324 DNA Homo sapiens anti-Rh(D) antibody clone SH32 209 gccgagctcactcaggagcc ctcactgact gtgtccccag gagggacagt cactctcacc 60 tgtgcttccagcactggagc agtcaccagt cgttactttc caaactggtt ccagcagaaa 120 cctggccaggcacccagggc actgatttat ggttcaaaca acaaacactc ctggacccct 180 gcccggttctcaggctccct ccttgggggc aaagctgccc tgacactgtc aggtgtgcag 240 cctgaggacgaggcggagta ttactgcctg ctcttctatg ctggtgcttg ggcgttcggc 300 ggagggaccaagctgaccgt ccta 324 210 324 DNA Homo sapiens anti-Rh(D) antibody cloneSH34 210 gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacagagtcaccatc 60 acttgccggg caagtcagag cattagcagc tatttaaatt ggtatcagcagaaaccaggg 120 aaagccccta agctcctgat ctatgctgca tccggtttgc aaagtggggtcccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtctgcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tacagtaccc ccccgtacacttttggccag 300 gggaccaagc tggagatcaa acga 324 211 321 DNA Homo sapiensanti-Rh(D) antibody clone SH36 211 gccgagctca ctcagtctcc atcctccctgtctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag cattagcagctatttaaatt ggtatcagca gaaaccaggg 120 aaatccccta agctcctgat ctatgctgcatccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg gacagatttcactctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactg tcaacagagttacagtaccc ctccggcttt cggccctggg 300 accaaagtgg atatcaaacg a 321 212 321DNA Homo sapiens anti-Rh(D) antibody clone SH39 212 gccgagctcacccagtctcc atcctccctg tctgcatctg tgggagacag agtcaccatc 60 acttgccgggcaagtcagac cattgggagg tatttaaatt ggtatcagca gaggccaggg 120 aaagcccccaaactcctggt atatgctgtg tccagtttgc aaagtggggc cccatcaagg 180 ttcagtggcagtggctctgg gacacatttc actctcacca tcaccagtct gcaacctgaa 240 gattttgcaacttacttctg ccaacagagt tacagttctc ctttcacttt tggccagggg 300 accaaggttgagatcaaacg a 321 213 321 DNA Homo sapiens anti-Rh(D) antibody clone SH41213 gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc 60acttgccggg caagtcagaa cattaggagg tctttaaatt ggtatcaaca caaaccaggg 120agagccccta gactcctgat ctatgctgca tccactttgc aaagtggggt cccatcaagg 180ttcaggggca gtggatctgg gacagatttc actctcacca tcaacagtct gcaacctgca 240gattttgcaa cttactactg tcagcagagt tccaataccc cgtggacgtt cggccatggg 300accaaggtgg aaatcaaacg a 321 214 318 DNA Homo sapiens anti-Rh(D) antibodyclone SH44 214 gccgagctca cccagtctcc atcgtccctg tctgcatctg taggagacagagtcatcatc 60 acttgccggg caagtcagac cattcccagg ttcttgaatt ggtatcaacagaagcctgga 120 aaagcccctg ttctcctgat tcatagtata tccagtttac aaagtggggtcccatcaagg 180 ttcagtgcca gtggatctgg gacagagttc actctcacca tcagcagtctgcaacctgaa 240 gatttcgcaa cttactactg ccaacagagt tacagtaatc tctctttcggccctgggacc 300 acagtggata ttagacga 318 215 321 DNA Homo sapiensanti-Rh(D) antibody clone SH46 215 gccgagctca cccagtctcc atcctccctgtctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagta cattagcagctatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta atctcctgat ctatgctgcatccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg gacagatttcactctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactg tcaacagacttacagttccc ctagcacttt cggccctggg 300 accaaagtgg atatcaaacg a 321 216 321DNA Homo sapiens anti-Rh(D) antibody clone SH47 216 gccgagctcacccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc 60 acttgccgggcaagtcagag cattagcaac tatttaaatt ggtatcagca gaaaccagga 120 aaagcccctaacctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggcagtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaacttactactg tcaacagagt tacagttatc ctcgcacgtt cggccaaggg 300 accaaggtggagatcagacg a 321 217 321 DNA Homo sapiens anti-Rh(D) antibody clone SH48217 gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc 60acttgccggg caagtcagta cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120aaagccccta atctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240gattttgcaa cttactactg tcaacagact tacagttccc ctagcacttt cggccctggg 300accaaagtgg atatcaaacg a 321 218 321 DNA Homo sapiens anti-Rh(D) antibodyclone SH49 218 gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacagagtcaccgtc 60 acttgccggg caagtcagag cattagcagc tatttaaatt ggtatcagcagaaaccaggg 120 aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggtcccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtctgcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tacagtaccc cgtggacgttcggccaaggg 300 accaaggtgg aaatcaaacg a 321 219 324 DNA Homo sapiensanti-Rh(D) antibody clone SH50 219 gccgagctca cccagtctcc atcgtccctgtctgcatctg taggagacag agtcaccatc 60 acttgccgga caagtcagag cattggcacctatttaaatt ggtatcaaca aaaaccaggg 120 aaagccccta aactcctgat ctatgctgcatccaatgtgc aaagtggggt cccatcaagg 180 ttcagtggcg gtggatctgg gacaggtttctctctcatca tcagcagtct gcaacctgaa 240 gatttagcaa tttactactg ccaacagagctacagtgtcc ctccgtacag ctttggcccg 300 gggaccaagc tggagatcaa acga 324 220324 DNA Homo sapiens anti-Rh(D) antibody clone SH51 220 gccgagctcacacagtctcc atccttcctg tctgcatctg taggagacag agtcaccatc 60 acttgccgggccagtcaggg cataaggagt tatttagcct ggtatcagca aaaaccaggg 120 aaagcccctaagctcctaat ctatgctgca tccactttgc aaagtggggt cccatcaagg 180 ttcagcggcagtggatctgg gacagaattc actctcacaa tcagcagcct gcagcctgaa 240 gattttgcaacttattactg tcaacagctt aataattacc cccctttcac tttcggccct 300 gggaccaaagtggatatcaa acga 324 221 321 DNA Homo sapiens anti-Rh(D) antibody cloneSH52 221 gccgagctca cccagtctcc atcctccatg tctgcatctg taggagacagagtcaccatc 60 acttgccggg caagtcagag cattggcact tatttaaatt ggtatcagcagaaaccaggg 120 aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggtcccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtctgcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tacagtaccc cgtggacgttcggccaaggg 300 accaaggtgg aaatcaaacg a 321 222 321 DNA Homo sapiensanti-Rh(D) antibody clone SH54 222 gccgagctca cccagtctcc atcctccatgtctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag cattggcacttatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta agctcctgat ctatgctgcatccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg gacagatttcactctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactg tcaacagagttacagtaccc cgtggacgtt cggccaaggg 300 accaaggtgg aaatcaaacg a 321 223 327DNA Homo sapiens anti-Rh(D) antibody clone SH55 223 gccgagctcacgcagccgcc ctcagcgtct gggacccccg ggcagagggt caccatctct 60 tgttctggaagcagctccaa catcggaagt aaatatgtat actggtacca gcaactccca 120 ggaacggcccccaaactcct catttatagt aataatcagc ggccctcagg ggtccctgac 180 cgattctctgccttcaagtc tggcacctca gcctccctgg ccatcactgg gctccaggct 240 gaggatgaggctaattatta ctgccagtcc tatgacagcg gcctgagtgg ctgggtgttc 300 ggcggcgggaccaagctgac cgtccta 327 224 324 DNA Homo sapiens anti-Rh(D) antibodyclone SH56 224 gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacagagtcaccatc 60 acttgccggg caagtcagag cattagcagg tatttaaatt ggtatcagcagaaaccaggg 120 aaagccccca agctcctgat ctatgctgca tccagtttgc aaagtggggtcccatcaagg 180 ttcagtggca gtggatctgg gacagatttc gctctcacca tcagcagtctgctacctgaa 240 gattttgcaa cttactactg tcaacagggt tacagtaccc ctccgtacagttttggccag 300 gggaccaagc tggagatcaa acga 324

What is claimed is:
 1. An isolated protein having an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 1-69 and 139-181.
 2. The isolated protein of claim 1, wherein the protein is an antigen-binding protein.
 3. The isolated protein of claim 2, wherein the antigen is human Rh(D) protein.
 4. The isolated protein of claim 1, wherein the protein has an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-69 and 139-181.
 5. The isolated protein of claim 1, wherein the protein is substantially purified.
 6. The isolated protein of claim 1, wherein the protein has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 1-27 and 139-153.
 7. The isolated protein of claim 6, wherein the protein has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 1-27.
 8. The isolated protein of claim 6, wherein the protein has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 139-153.
 9. The isolated protein of claim 1, wherein the protein has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 28-69 and 154-181.
 10. The isolated protein of claim 9, wherein the protein has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 28-44.
 11. The isolated protein of claim 9, wherein the protein has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 45-61.
 12. The isolated protein of claim 9, wherein the protein has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 62-69, and 154-163.
 13. The isolated protein of claim 9, wherein the protein has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 164-181.
 14. The isolated protein of claim 4, wherein the protein has a sequence selected from the group consisting of SEQ ID NOs: 1-27 and 139-153.
 15. The isolated protein of claim 14, wherein the protein has a sequence selected from the group consisting of SEQ ID NOs: 1-27.
 16. The isolated protein of claim 14, wherein the protein has a sequence selected from the group consisting of SEQ ID NOs: 139-153.
 17. The isolated protein of claim 4, wherein the protein has a sequence selected from the group consisting of SEQ ID NOs: 28-69 and 154-181.
 18. The isolated protein of claim 17, wherein the protein has a sequence selected from the group consisting of SEQ ID NOs: 28-44.
 19. The isolated protein of claim 17, wherein the protein has a sequence selected from the group consisting of SEQ ID NOs: 45-61.
 20. The isolated protein of claim 17, wherein the protein has a sequence selected from the group consisting of SEQ ID NOs: 62-69, and 154-163.
 21. The isolated protein of claim 17, wherein the protein has a sequence selected from the group consisting of SEQ ID NOs: 164-181. 